响应土壤阴离子类型的盐碱土古细菌群落多样性研究

为初步解析我国黑龙江苏打盐碱地（HA）、新疆荒漠盐碱地（XD）、山西平原盐碱地（SY）、江苏滨海盐碱地（JD）和天津滨海盐碱地（TB）5个地区的土壤古细菌群落结构多样性,以及其响应土壤不同阴离子类型的分布特征,采用基于Illumin-Hiseq平台的高通量测序技术获得489~604个古细菌可操纵分类单元（OTU）.OTU多样性分析结果表明XD的古细菌群落物种丰富度最高,SY和HA分别具有最高和最低的古细菌群落多样性和均匀度;OTU的物种注释结果表明HA以奇古菌门（Thaumarchaeota）为优势菌门,其他土壤均以广古菌（Euryarchaeota）为优势;综合5个样品古细菌群落结构、优势古细菌属和土壤阴离子进行的典范对应分析（CCA）结果显示,XD古细菌群落结构和Haloterrigena属（XD中最优势,相对丰度20.2%）明显的响应土壤SO₄^2-浓度;HA古细菌群落结构和Nitrososphaera属（HA中最优势,62.3%）明显的响应土壤HCO₃^-/CO₃^2-浓度和pH;JD、SY和TB古细菌群落结构、Halorubrum属（JD和TB中最优势,24.4%和15.6%）,和Natronomonas属（SY和JD中第二优势,8.1%和9.2%）明显的响应土壤Cl^-浓度.研究结果说明我国不同地域分布的盐碱土受其土壤阴离子类型及其浓度的影响,呈现不同的古细菌群落结构和分布特征.研究结果为我国不同类型盐碱土古细菌资源的挖掘提供依据,也为揭示我国不同类型盐碱土古细菌群落的生态功能提供参考.     

Effect of reclaimed water effluent on bacterial community structure in the Typha angustifolia L. rhizosphere soil of urbanized riverside wetland, China

In order to evaluate the impact of reclaimed water on the ecology of bacterial communities in the Typha angustifolia L. rhizosphere soil,bacterial community structure was investigated using a combination of terminal restriction fragment length polymorphism and 16S rRNA gene clone library. The results revealed significant spatial variation of bacterial communities along the river from upstream and downstream. For example,a higher relative abundance of γ-Proteobacteria,Firmicutes,Chloroflexi and a lower proportion ofβ-Proteobacteria and ε-Proteobacteria was detected at the downstream site compared to the upstream site. Additionally,with an increase of the reclaimed water interference intensity,the rhizosphere bacterial community showed a decrease in taxon richness,evenness and diversity. The relative abundance of bacteria closely related to the resistant of heavy-metal was markedly increased,while the bacteria related for carbon/nitrogen/phosphorus/sulfur cycling wasn't strikingly changed. Besides that,the pathogenic bacteria markedly increased in the downstream rhizosphere soil since reclaimed water supplement,while the possible plant growth-promoting rhizobacteria obviously reduced in the downstream sediment.Together these data suggest cause and effect between reclaimed water input into the wetland,shift in bacterial communities through habitat change,and alteration of capacity for biogeochemical cycling of contaminants.     

Screening and degrading characteristics and community structure of a high molecular weight polycyclic aromatic hydrocarbon-degrading bacterial consortium from contaminated soil

Inoculation with effcient microbes had been proved to be the most important way for the bioremediation of polluted environments. For the treatment of abandoned site of Beijing Coking Chemical Plant contaminated with high level of high-molecular-weight polycyclic aromatic hydrocarbons(HMW-PAHs) ,a bacterial consortium capable of degrading HMW-PAHs,designated 1-18-1,was enriched and screened from HMW-PAHs contaminated soil.Its degrading ability was analyzed by high performance liquid chromatography(HPLC) ,and the community structure was investigated by construction and analyses of the 16S rRNA gene clone libraries(A,B and F) at different transfers.The results indicated that 1-18-1 was able to utilize pyrene,fluoranthene and benzoapyrene as sole carbon and energy source for growth.The degradation rate of pyrene and fluoranthene reached 82.8%and 96.2%after incubation for 8 days at 30°C,respectively;while the degradation rate of benzoapyrene was only 65.1%after incubation for 28 days at 30°C. Totally,108,100 and 100 valid clones were randomly selected and sequenced from the libraries A,B,and F.Phylogenetic analyses showed that all the clones could be divided into 5 groups,Bacteroidetes,α-Proteobacteria,Actinobacteria,β-Proteobacteria andγ-Proteobacteria.Sequence similarity analyses showed total 39 operational taxonomic units(OTUs) in the libraries.The predominant bacterial groups wereα-Proteobacteria(19 OTUs,48.7%) ,γ-Proteobacteria(9 OTUs,23.1%) andβ-Proteobacteria(8 OTUs,20.5%) . During the transfer process,the proportions ofα-Proteobacteria andβ-Proteobacteria increased greatly(from 47%to 93%) ,while γ-Proteobacteria decreased from 32%(library A) to 6%(library F) ;and Bacteroidetes group disappeared in libraries B and F.     

Analysis of bacterial community in bulking sludge using culture-dependent and -independent approaches

Combination of 16S rRNA gene clone library and cultivation for assessing of bacterial diversity of the microflora in bulking sludge was evaluated in the study.     

长江口潮滩沉积物古菌群落结构与多样性

以长江口潮滩为研究对象,采用高通量测序技术,研究了潮滩沉积物古菌群落结构与多样性及其影响因素.结果表明,沉积物中古菌群落OTUs数量为900~1417,Shannon指数为7.02~8.02,均表现出从低盐度向高盐度样点降低的变化特征.各采样点古菌群落特异性OTUs占各采样点总OTUs的24.2%~57.3%,而共有的OTUs仅占1.2%,说明各样点沉积物中古菌群落特异性较高.沉积物中古菌主要隶属于广古菌门（Euryarchaeota）和奇古菌门（Thaumarchaeota）,而深古菌门（Bathyarchaeota）也是古菌群落的重要组成部分,其相对丰度占到17.7%~25.9%.主成分和聚类分析表明芦潮港和东海农场沉积物中古菌群落结构相似,白龙港和浏河口群落组成相近,而浒浦与其他采样点的古菌群落组成差别较大.通过典范对应分析,研究区沉积物中古菌群落结构与沉积物盐度水平有着非常密切的关系.上述研究结果表明长江口潮滩沉积物中古菌群落结构组成和多样性具有高度的空间差异性,且沉积物盐度是群落差异的决定性影响因素.     

Soil fungistasis and its relations to soil microbial composition and diversity: a case study of a series of soils with different fungistasis

Fungistasis is one of the important approaches to control soil-borne plant pathogens.Some hypotheses about the mechanisms for soil fungistasis had been established,which mainly focused on the soil bacterial community composition,structure,diversity as well as function.In this study,the bacterial community composition and diversity of a series of soils treated by autoclaving,which coming from the same original soil sample and showing gradient fungistasis to the target soil-borne pathogen fungi Fusarium gr...     

玉米秸秆厌氧降解复合菌系的微生物群落结构

以不同来源具有木质纤维素降解能力的环境样品为接种物,以玉米秸秆为唯一碳源,进行了秸秆厌氧降解微生物的驯化培养,构建了8组复合菌系.复合菌系的产气周期为30～50 d,玉米秸秆的总固体(TS)去除率在30%～40%之间.其中6组的甲烷产量(以TS计)为62.1～118.4 mL.g-1,发酵液中挥发性有机酸主要为乙酸、丙酸及丁酸(100～500 mg.L-1),发酵终产物pH为6.5～6.7;而另外2组的甲烷产量较低,在8.5～9.7 mL.g-1之间,但乙酸浓度较高(1 200 mg.L-1左右),发酵终产物pH为5.6～5.9.通过16S rRNA基因克隆文库方法对复合菌系中细菌及古菌的群落结构进行了多样性解析,结果表明细菌主要分为8个不同的类群,其中厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)、互养菌门(Synergistetes)及热袍菌门(Thermotogae)为优势菌群,分别占细菌克隆总数的37.8%、34.3%、11.6%和6.4%;古菌属于甲烷微菌纲(Methanomicrobia)和甲烷杆菌纲(Methanobacteria),分别占古菌克隆总数的61.1%和38.9%.对主要功能菌群在秸秆厌氧降解产甲烷过程中发挥的作用进行了探讨.     

Soil fungistasis and its relations to soil microbial composition and diversity: A case study of a series of soils with di erent fungistasis

Fungistasis is one of the important approaches to control soil-borne plant pathogens.Some hypotheses about the mechanisms for soil fungistasis had been established,which mainly focused on the soil bacterial community composition,structure,diversity as well as function.In this study,the bacterial community composition and diversity of a series of soils treated by autoclaving,which coming from the same original soil sample and showing gradient fungistasis to the target soil-borne pathogen fungi Fusarium gr...     

Eco-toxicity of petroleum hydrocarbon contaminated soil

Total petroleum hydrocarbons (TPH) contaminated soil samples were collected from Shengli Oilfield of China. Toxicity analysis was carried out based on earthworm acute toxicity, plant growth experiment and luminescent bacteria test. The soil was contaminated bypetroleum hydrogcarbons with TPH concentration of 10.57%. With lethal and sub-lethal rate as endpoint, earthworm test showed that the LD50 (lethal dose 50%) values in 4 and 7 days were 1.45% and 1.37% respectively, and the inhibition rate of earthworm body weight increased with higher oil concentration. TPH pollution in the soil inhibited seed germination in both wheat and maize experiment when the concentration of petroleum was higher than 0.1%. The EC50 (e ective concentration 50%) for germination is 3.04% and 2.86% in maize and wheat, respectively. While lower value of EC50 for root elongation was to be 1.11% and 1.64% in maize and wheat, respectively, suggesting higher sensitivity of root elongation on petroleum contamination in the soil. The EC50 value in luminescent bacteria test was 0.47% for petroleum in the contaminated soil. From the experiment result, it was concluded that TPH content of 1.5% is considered to be a critical value for plant growth and living of earthworm and 0.5% will a ect the activity of luminescent bacteria.     

同时产甲烷反硝化颗粒污泥中微生物群落结构

在UASB成功运行同时产甲烷反硝化小试基础上,针对反应器内颗粒污泥,通过构建古菌和细菌的16SrDNA基因文库、RADAR遗传变异分型和测序比对等技术进行了微生物系统发育关系和群落结构分析.结果表明,在颗粒污泥的古菌中,产甲烷髦毛菌和产甲烷杆菌是主要菌群,随机选出的88个古菌克隆子,这两类古菌的16SrDNA序列分别占古菌总量71.59%和22.73%;而颗粒污泥中细菌的多样性要高于古菌,低GC革兰氏阳性菌和ε变型菌纲分支的细菌是细菌的主要菌群,在随机选出的133个细菌克隆子中,这2类细菌的16SrDNA序列分别占细菌总量的49.62%和12.03%.     

8种花卉植物种子萌发对石油烃污染土壤的响应

石油污染土壤的植物修复技术以其处理成本低、无二次污染、自然美观等特点,正逐步成为石油污染治理研究的一个重要方向.而修复植物的筛选是植物修复技术首要解决的问题.为此以花卉植物种子发芽作为生态指示,探讨其对石油污染土壤的反应.设置了4组不同浓度的石油污染土壤处理,对8种供试花卉植物的种子发芽率进行了盆栽实验观测.结果表明:不同的花卉种子对石油污染表现出不同的耐受性,与清洁对照土壤相比,大多数花卉种子的萌发都明显受到石油烃污染的抑制,其中受石油烃污染影响最严重的是二月兰(Orychophragmus violace).相比之下,4组浓度处理下硫华菊(Cosmos sulphnreus)和孔雀草(Tagetes patula)具有较高的耐受性和种子发芽率,因而具有修复石油污染土壤的潜力.     

新型生态浮岛在改善水质中的作用及生物膜载体微生物特征研究

设计了一种新型生态浮岛(IEFI),并对浮岛生物膜载体上的微生物进行了研究.结果表明,生物膜载体上的微生物对改善水质起到重要的作用,新型生态浮岛对COD、TN、TP的去除效果明显高于普通浮岛(CFI)(P<0.05).COD、TN、TP的最高去除率分别出现在9月、5月和11月.构建了生物膜载体上微生物的克隆文库,结果表...     

德国分散式污水处理系统微生物群落结构及功能

以德国分散式污水处理系统为研究对象,通过16S rRNA测序,研究2个反应器中微生物群落结构,利用PICRUSt软件对其功能进行推演.结果表明,冬季,污水厌氧膜生物反应器（AnMBR）内温度20℃,进水COD 712mg/L时,出水可获得52%的COD平均去除率,产气率为122L/kgCOD;固体废物厌氧反应器（PSD）内温度37℃,反应器内COD 3007mg/L时,可获得374L/kgVSS的产气率.2个反应器具有相似的微生物组成,对细菌,Synerigistaceae科的相对丰度最高（AnMBR：24.0%±10.0%;PSD：11.0%±3.1%）;对古菌,Methanobacteriaceae科的相对丰度最高（AnMBR：0.6%±0.3%;PSD：13.8%±1.8%）;2个反应器的功能基因组成也相似,产甲烷都以H₂还原CO₂的通路为主.PSD反应器中H₂还原CO₂通路相关基因、F420合成相关基因、辅酶M合成相关基因的相对丰度都高于AnMBR反应器.     

Assessing the impact of fungicide enostroburin application on bacterial community in wheat phyllosphere

Fungicides have been used extensively for controlling fungal pathogens of plants. However, little is known regarding the effects that fungicides upon the indigenous bacterial communities within the plant phyllosphere. The aims of this study were to assess the impact of fungicide enostroburin upon bacterial communities in wheat phyllosphere. Culture-independent methodologies of 16S rDNA clone library and 16S rDNA directed polymerase chain reaction with denaturing gradient gel electrophoresis (PCR-DGGE) were used for monitoring the change of bacterial community. The 16S rDNA clone library and PCR-DGGE analysis both confirmed the microbial community of wheat plant phyllosphere were predominantly of the γ-Proteobacteria phyla. Results from PCR-DGGE analysis indicated a significant change in bacterial community structure within the phyllosphere following fungicide enostroburin application. Bands sequenced within control cultures were predominantly of Pseudomonas genus, but those bands sequenced in the treated samples were predominantly strains of Pantoea genus and Pseudomonas genus. Of interest was the appearance of two DGGE bands following fungicide treatment, one of which had sequence similarities (98%) to Pantoea sp. which might be a competitor of plant pathogens. This study revealed the wheat phyllosphere bacterial community composition and a shift in the bacterial community following fungicide enostroburin application.     

石油污染土壤热解修复反应规律与机制研究

在石油的开采、运输等过程中,由于原油泄漏而产生的石油污染土壤会严重威胁生态环境和人类健康.相比细菌降解修复和化学氧化修复,中低温热解修复是一种高效、经济的修复方法.然而,国内对石油污染土壤的热解修复规律和影响因素研究却较少.本文先以模型污染土壤为研究对象,考察了初始含油率(5％、10％)、热解温度(375、400、42...     

Microbial changes in rhizospheric soils contaminated with petroleum hydrocarbons after bioremediation

Effects of bioremediation on microbial communities jn soils contaminated with petroleum hydrocarbons are a scientific problem to be solved. Changes in dominate microbial species and the total amount of microorganisms including bacteria and fungi in rhizospheric soils after bioremediation were thus evaluated using field bioremediation experiments. The results showed that there were changed dominant microorganisms including ] ] bacterial strains which are mostly Gram positive bacteria and 6 fungal species which were identified. The total amount of microorganisms including bacteria and fungi increased after bioremediation of microbial agents combined with planting maize. On the contrary, fungi in rhizospheric soils were inhibited by adding microbial agents combined with planting soybean.     

某石油污染地下水溶解性无机碳低异常的微生物地球化学成因探析

微生物地球化学作用往往导致石油污染场地地下水中溶解性无机碳(DIC)升高,而华北某石油污染场地地下水DIC低异常明显.为究其机理,在水文地球化学分析和16S rRNA基因高通量测序基础上,结合含水层结构及流场特征,剖析了地下水水化学和微生物两方面作用,辨识了地下水中DIC变异的主导因素,揭示了其中的生物地球化学作用的机理,发现该场地地下水DIC低异常可能与地下水中具有较高浓度钙镁离子和较高活性自养微生物有关,自养微生物代谢及诱导产生碳酸盐岩沉淀作用极可能是该场地地下水中DIC低异常的成因.推测机理为:Hydrogenophaga和Sedimentibacter等菌属微生物在氢化酶的作用下产氢气,Hydrogenophaga、Pseudomonas、Pseudoxanthomonas、Polynucleobacter等固碳微生物和产甲烷微生物利用氢气作为能源,将DIC合成有机碳,并产生碱性微环境,促使Ca~(2+)和Mg~(2+)与DIC反应形成碳酸钙镁沉淀.     

高通量测序技术应用于污水处理厂细菌气溶胶群落结构分析

由于污水及其处理设施中细菌气溶胶的逸散作用,导致污水处理厂细菌气溶胶的群落结构较为复杂,因此,选择适宜的采样和分析方法对其多样性的认识和风险评估具有重要意义.本研究以某城市污水厂为研究对象,采用"总悬浮颗粒物采样器+高通量测序技术"分别对污水处理全过程进行细菌气溶胶的收集与分析;作为对照,采用"Andersen六级采样器+克隆文库技术"进行同步采样和分析.结果表明,前者对各采样点气溶胶中细菌的多样性、总体群落组成、优势群落的解析均明显优于后者,分析更简便快捷,数据更全面、更接近其真实的分布状态.本研究结果可为污水处理厂细菌气溶胶的采样和分析方法的选择提供科学依据.     

Vertical response of microbial community and degrading genes to petroleum hydrocarbon contamination in saline alkaline soil

A column microcosm was conducted by amending crude oil into Dagang Oilfield soil to simulate the bioremediation process. The dynamic change of microbial communities and metabolic genes in vertical depth soil from 0 to 80 cm were characterized to evaluate the petroleum degradation potential of indigenous microorganism. The influence of environmental variables on the microbial responds to petroleum contamination were analyzed. Degradation extent of 42.45% of n-alkanes(C8–C40) and 34.61% of 16ΣPAH were reached after 22 weeks. Relative abundance of alkB, nah, and phe gene showed about 10-fold increment in different depth of soil layers. Result of HTS profiles demonstrated that Pseudomonas, Marinobacter and Lactococcus were the major petroleum-degrading bacteria in0–30 and 30–60 cm depth of soils. Fusarium and Aspergillus were the dominant oil-degrading fungi in the 0–60 cm depth of soils. In 60–80 cm deep soil, anaerobic bacteria such as Bacteroidetes, Lactococcus, and Alcanivorax played important roles in petroleum degradation.Redundancy analysis(RDA) and correlation analysis demonstrated that petroleum hydrocarbons(PHs) as well as soil salinity, clay content, and anaerobic conditions were the dominant effect factors on microbial community compositions in 0–30, 30–60, and 60–80 cm depth of soils, respectively.     

Bacterial diversity and distribution in the southeast edge of the Tengger Desert and their correlation with soil enzyme activities

The nature of microbial communities and their relation to enzyme activities in desert soils is a neglected area of investigation. To address this, the bacterial diversity and distribution and soil physico-chemical factors were investigated in the soil crust, the soil beneath the crust and rhizosphere soil at the southeast edge of the Tengger Desert, using the denaturing gradient gel electrophoresis of 16S rRNA genes amplified by the polymerase chain reaction. Phylogenetic analysis of the sequenced DGGE bands revealed a great diversity of bacteria. The Proteobacteria, consisting of the α, β, and γ subdivisions, were clearly the dominant group at all depths and in rhizosphere soil. Analysis of the enzyme activities indicated that the rhizosphere soil of Caragana korshinskii exhibited the highest protease and polyphenol oxidase activities, and in the soil crust there were increased activities of catalase, urease, dehydrogenase and sucrase. The bacterial community abundance closely correlated with soil enzyme activities in different soils. The presence of Cyanobacteria correlated with significant increases in protease, catalase and sucrase in the soil crust, and increased urease in the rhizosphere soil of Artemisia ordosica. The occurrence of Acidobacteria was associated with significant increases in urease, dehydrogenase, and sucrase in the rhizosphere soil of C. korshinski. The presence of γ-Proteobacteria correlated with a significant increase in polyphenol oxidase in the rhizosphere soil of A. ordosica. The study indicated a close relationship between the soil bacterial community and soil enzymes, suggesting the necessity of further investigations into bacterial function in this desert ecosystem.