株洲清水塘工业区周边土壤微生物群落特征

基于Illumina高通量测序技术分析了株洲清水塘工业区周边土壤的微生物群落特征,研究重金属污染对土壤微生物群落的影响.结果表明,土壤微生物群落的相对丰度和多样性变化趋势一致,均随着重金属污染程度增加而减小;稻田土壤平均相对丰度最高的门是变形菌门(49. 56%),其次为绿弯菌门(13. 07%)和酸杆菌门(8. 77%);较高重金属污染程度下伴随着更高丰度的变形菌门、绿弯菌门与更低丰度的硝化螺旋菌门、酸杆菌门; 4组样品的微生物群落结构相似性较高,OTU重叠度达52. 64%,丰度较高的OTU其微生物群落更趋向于相似;结合Spearman相关性分析,Cr与变形菌门显著负相关,与绿弯菌门显著正相关,Cd、Cu、Pb和Zn与硝化螺旋菌门显著负相关.以上结果表明,重金属污染是影响清水塘工业区周边土壤微生物群落结构的重要因素.     

堆肥-生物强化对重度石油污染土壤的修复作用

为解决微生物强化修复油污土壤过程中降解菌在低温环境下活性较低的问题,利用有机堆肥作为固体培养基对降解菌进行扩大培养,将获得的降解菌-堆肥制剂施入油污土壤中进行修复研究.利用重量法和GC-MS分析土壤中石油烃含量变化,利用Illumina Mi Seq对土壤微生物群落结构进行分析.结果表明,利用堆肥作为固体基质可对降解菌进行扩大培养.低温环境下利用堆肥-降解菌制剂对油污土壤修复30 d,土壤中石油烃、烷烃、多环芳烃去除率分别为27.0%、19.6%、10.0%;自然放置的土壤中3种烃去除率分别为4.5%、9.5%、2.3%.加入降解菌-堆肥制剂进行修复的土壤香农指数和Ace指数分别由4.42和1718.5增加为5.30和2170.5;土壤中变形菌门(Proteobacteria)和放线菌门(Actinobacteria)所占丰度由53.4%和25.9%分别降低至48.9%和14.1%,拟杆菌门(Bacteroidetes)所占丰度由5.0%增加至24.5%.属水平上,不动细菌属(Acinetobacter)和假单胞菌属(Pseudomonas)所占丰度由0.02%和3.4%分别提高至15.2%和4.6%.研究结果表明,在低温条件下向石油污染土壤中施入降解菌-堆肥制剂可提高土壤中的石油烃去除率,并使土壤微生物群落结构发生明显变化.     

微生物修复油污土壤过程中氮素的变化及菌群生态效应

对陕北地区石油污染土壤进行了微生物修复研究,利用气相色谱-质谱联用技术(gas chromatography-mass spectrometry,GC-MS)测定土壤中烷烃和多环芳烃的含量变化,采用高通量测序技术研究微生物修复对油污土壤微生物群落多样性的影响,对修复过程中土壤不同形态氮含量变化进行了分析测定.结果表明,经过18周的修复,土壤中烷烃含量由初始时的25 987.8mg·kg~(-1)降低为12 788.6 mg·kg~(-1),多环芳烃含量由初始时的5 322.9 mg·kg~(-1)降低为2 917.2mg·kg~(-1).高通量测序结果表明,经过微生物修复处理的土壤微生物群落结构发生了较大变化,一些可降解烃类的菌群如厚壁菌门(Firmicutes)、拟杆菌门(Bacterodetes)、迪茨氏菌属(Dietzia)、不动杆菌属(Acinetobacter)所占丰度增加.经过修复处理的土壤中总氮、氨氮含量呈降低趋势,硝态氮含量在修复前期呈降低趋势,修复后期基本保持不变.研究结果表明,微生物修复处理可去除土壤中的大部分烷烃和多环芳烃,烷烃和多环芳烃的降解效果与土壤中硝氮含量和烃降解菌所占丰度相关.     

基于高通量测序分析的生物修复石油污染土壤菌群结构变化

利用高通量测序技术对微生物修复石油污染土壤过程中的微生物群落结构变化进行研究.结果表明,经修复处理的土壤微生物群落结构及多样性发生明显变化.利用生物强化修复处理(BA)的土壤中,微生物丰富度与均匀度明显降低,土著菌群受到抑制,外加变形菌门(Proteobacteria)成为主要的优势菌门,相对丰度由修复前的37. 44%增加为87. 44%.假单胞菌属(Pseudomonas)成为土壤中的优势菌属,丰度由2. 99%增加为76. 37%;进行生物刺激修复处理的土壤(BS)菌群丰富度和均匀度与原污染土壤相比略有降低.菌群结构组成上,原优势菌门变形菌门(Proteobacteria)丰度由37. 44%降低为10. 90%,厚壁菌门(Firmicutes)丰度由9. 16%增加为35. 32%,属水平上,原优势菌属微小杆菌属(Exiguobacterium)和原小单胞菌属(Promicromonospora)丰度由8. 49%和18. 96%分别降低为2. 19%和14. 97%,诺卡氏菌属(Nocardioides)和芽孢杆菌属(Bacillus)丰度由5. 56%和0. 29%分别增加至28. 95%和22. 70%,成为主要优势菌属.生物强化修复处理引起土壤菌群多样性和结构发生明显变化,生物刺激修复处理可基本保持土壤菌群结构多样性不被破坏,土壤菌群结构的稳定有利于石油烃的生物降解.     

基于高通量测序分析的生物修复石油污染土壤菌群结构变化

利用高通量测序技术对微生物修复石油污染土壤过程中的微生物群落结构变化进行研究。结果表明,经修复处理的土壤微生物群落结构及多样性发生明显变化。利用生物强化修复处理(BA)的土壤中,微生物丰富度与均匀度明显降低,土著菌群受到抑制,外加变形菌门(Proteobacteria)成为主要的优势菌门,相对丰度由修复前的37.44%增加为87.44%。假单胞菌属(Pseudomonas)成为土壤中的优势菌属,丰度由2.99%增加为76.37%;进行生物刺激修复处理的土壤(BS)菌群丰富度和均匀度与原污染土壤相比略有降低。菌群结构组成上,原优势菌门变形菌门(Proteobacteria)丰度由37.44%降低为10.90%,厚壁菌门(Firmicutes)丰度由9.16%增加为35.32%,属水平上,原优势菌属微小杆菌属(Exiguobacterium)和原小单胞菌属(Promicromonospora)丰度由8.49%和18.96%分别降低为2.19%和14.97%,诺卡氏菌属(Nocardioides)和芽孢杆菌属(Bacillus)丰度由5.56%和0.29%分别增加至28.95和22.70%,成为主要优势菌属。生物强化修复处理引起土壤菌群多样性和结构发生明显变化,生物刺激修复处理可基本保持土壤菌群结构多样性不被破坏,土壤菌群结构的稳定有利于石油烃的生物降解。     

石油污染土壤富集前后细菌群落组成和共现网络分析

为了探究石油污染土壤中细菌群落在富集过程中的演替规律,试验采用平板划线法、菌落PCR和高通量测序技术,分析了富集前后细菌群落结构、共现网络和核心菌属组成,并对富集后体系中的微生物进行分离鉴定,筛选石油降解菌.研究表明富集体系中可培养微生物隶属于34个属53个种,其中3个为潜在新种微生物,Dietzia maris OS33和Rhodococcus qingshengii OS62-1具有降解石油的能力.高通量测序结果显示,在门分类水平上,富集前后丰度较高的菌门均为Proteobacteria和Actinobacteria,富集后两菌门的丰度可达到97.98%,占据绝对优势;丰度较高菌属由Pseudomonas、Rhodococcus、Bacillus和Xanthomonas转变为Dietzia、Unspecified_Idiomarianceae和Halomonas,标志微生物转变为与石油降解有关的Dietzia.细菌群落共现网络在富集后,网络结构进一步简化且更加稳定,核心微生物转变为与石油降解有关的Pseudomonas、Lysinibacillus、Pseudochrobactrum、Agrobacterium、Lactobacillus,且非石油降解菌P.songnenensis P35可协同石油降解菌D.maris OS33降解石油.     

邻苯二甲酸酯对土壤微生物生态质量的影响研究

文章从微生物群落差异角度研究了不同浓度的邻苯二甲酸酯(PAEs)对土壤质量的影响。结果表明:随着PAEs浓度的增加,变形菌门Proteobacteria、厚壁菌门Firmicutes的相对丰度降低,而酸杆菌门Acidobacteria、拟杆菌门Bacteroidetes和绿弯菌Chloroflexi等菌门的相对丰度增加;Acidobacteria_Gp6、Acidobacteria_Gp4、Acidobacteria_Gp7等多种不动杆菌,假单胞菌属Pseudomonas,戈登氏菌属Gordonia和芽孢杆菌属Bacillus等常被报道具有降解PAEs功能菌属在样品中所占的丰度也随着PAEs的浓度增加而大幅上升;PAEs污染降低土壤中细菌群落的多样性,从而降低土壤的生态质量。     

不同淋洗剂淋洗对镉污染农田土壤微生物群落结构的影响

土壤微生物群落对重金属污染土壤修复效果有较好的响应.为评价不同淋洗剂淋洗对镉（Cd）污染农田土壤微生物群落结构的影响,选用Cd污染土壤常用的乙二胺四亚甲基膦酸（EDTMP）和乙二胺四乙酸（EDTA）作为供试淋洗剂,对采自矿区周边Cd重度污染农田土壤进行室内浸提试验,分别测定了浸提后土壤微生物群落结构、Cd的化学形态和部分理化性质.结果表明,EDTMP和EDTA浸提对土壤Cd都具有一定的去除效果,其单次浸提率分别为14.23%和38.93%.浸提后土壤优势菌群结构发生明显变化,经EDTMP浸提土壤变形菌门相对丰度显著提高,而土壤酸杆菌门和硝化螺旋菌门的相对丰度则显著降低;经EDTA浸提土壤拟杆菌门相对丰度显著降低.经EDTMP浸提土壤的变形菌门、拟杆菌门和浮霉菌门的相对丰度显著高于经EDTA浸提土壤,芽单胞菌门和硝化螺旋菌门的相对丰度则显著低于经EDTA浸提后的土壤（p<0.05）.冗余分析（RDA）结果表明,2种淋洗剂主要通过降低土壤Cd含量、改变土壤pH及主要养分含量来影响微生物群落结构.     

石油污染对土壤微生物群落影响及石油降解菌的筛选鉴定

为研究石油污染对土壤中细菌群落结构及土壤理化性质的影响,分离筛选石油降解菌,从陕北宝塔、吴起、靖边和延长4个县区采集石油污染土壤和未受石油污染土壤,测量土壤中石油、有机质、硝态氮、铵态氮、速效磷、速效钾的含量以及pH;采用高通量测序技术对2种土壤中细菌群落结构进行比较分析;并以石油为唯一碳源,从石油污染土壤中筛选高效石油降解菌,并对所筛选的高效石油降解菌进行16S rDNA鉴定。陕北4个县区石油污染土壤中铵态氮、硝态氮、速效磷和速效钾含量分别降低了0.57,6.63,4.34,8.91 mg/kg,有机质含量增加了2~21倍。石油污染土壤中细菌群落的丰富度和多样性均降低,其中变形菌门(Proteobacteria)和绿弯菌门(Chloroflexi)为主要菌门,分枝杆菌属(Mycobacterium)为丰度最高菌属。以石油为唯一碳源,分离得到8株石油降解菌,其中菌株OS33和菌株OS62-1在5 d内的石油降解率分别为80.51%和81.60%,经鉴定OS33为迪茨氏菌(Dietzia sp.),OS62-1为红球菌(Rhodococcus sp.)。石油污染发生后,土壤中细菌群落的丰富度和多样性降低,筛选的8株石油降解菌中OS62-1石油降解率最高,研究结果进一步丰富了陕北地区石油降解菌菌种资源库。     

泗顶矿区6种土地利用类型土壤微生物群落结构特征

以广西柳州泗顶铅锌矿区的玉米地、菜地、灌木、尾矿库、森林和柑橘园6种不同利用类型土壤为研究对象,采用高通量测序技术,分析了不同土地利用类型土壤微生物群落结构特征及土壤环境因子对微生物群落结构的影响.结果表明,变形菌门（Proteobacteria）是6种土壤的优势菌门,所占比例为31.34%~53.47%,γ-变形菌纲（Gammaproteobacteria）为优势菌纲.冗余分析表明,在重金属含量最高的灌木区,厚壁菌门（Firmicutes）和拟杆菌门（Bacteroidetes）的丰度与土壤中铅、锌、镉、铜的含量呈现显著正相关;在土壤钠和钾含量较高的玉米地和菜地,浮霉菌门（Planctomycetes）的丰度有所增加,而厚壁菌门和拟杆菌门的丰度则有所下降.在土壤有机质含量较高而总氮、总磷含量较低的柑橘园,酸杆菌门（Acidobacteria）和疣微菌门（Verrucomicrobia）的丰度较高.芽单胞菌门（Gemmatimonadetes）和浮霉菌门的丰度与土壤pH值呈显著正相关.研究结果表明土地利用类型及土壤环境因子对土壤微生物群落结构多样性有显著的影响.     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.     

Removal of heavy metals from sewage sludge by low costing chemical method and recycling in agriculture

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Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.     

Single and joint effects of pesticides and mercury on soil urease

The influence of two pesticides including chlorimuron-ethyl and furadan and mercury （Hg） on urease activity in 4 soils （meadow burozem and phaeozem） was investigated. The soils were exposed to various concentrations of the two pesticides and Hg individually and simultaneously. Results showed that there was a close relationship between urease activity and organic matter content in soil. Chlorimuron-ethyl and furadan could both activate urease in the 4 soils. The maximum increment of urease activity by chlorimuronethyl was up to 14%-18%. There was almost an equal increase （up to 13%-21%） in the urease activity by furadan. On the contrary, Hg markedly inhibited soil urease activity. A logarithmic equation was used to describe the relationship （P〈0.05） between the concentration of Hg and the activity of soil urease in the 4 tested soils. Semi-effect dose （ED50） values by the stress of Hg based on the inhibition of soil urease in the 4 soils were 88, 5.5, 24 and 20 mg/kg, respectively, according to the calculation of the corresponding equations. The interactive effect of chlorimuron-ethyl or furadan with metal Hg on soil urease was mainly synergic at the highest tested concentrations.     

Organochlorine pesticides in soils under different land usage in the Taihu Lake region, China

A field study was conducted in the Taihu Lake region, China in 2004 to reveal the organochlorine pesticide concentrations in soils after the ban of these substances in the year 1983. Thirteen organochlorine pesticides (OCPs) were analyzed in soils from paddy field, tree land and fallow land. Total organochlorine pesticide residues were higher in agricultural soils than in uncultivated fallow land soils. Among all the pesticides, ΣDDX (DDD, DDE and DDT) had the highest concentration for all the soil samples, ranging from 3.10 ng/g to 166.55 ng/g with a mean value of 57.04 ng/g and followed by ΣHCH, ranging from 0.73 ng/g to 60.97 ng/g with a mean value of 24.06 ng/g. Dieldrin, endrin, HCB and α-endosulfan were also found in soils with less than 15 ng/g. Ratios of p,p'-(DDD DDE)/DDT in soils under three land usages were: paddy field ＞ tree land ＞ fallow land, indicating that land usage inlfuenced the degradation of DDT in soils. Ratios of p,p'-(DDD DDE)/DDT ＞1, showing aged residues of DDTs in soils of the Taihu Lake region. The results were discussed with data from a former study that showed very low actual concentrations of HCH and DDT in soils in the Taihu Lake region, but according to the chemical half-lives and their concentrations in soils in 1980s, the concentration of DDT in soils seemed to be underestimated. In any case our data show that the ban on the use of HCH and DDT resulted in a tremendous reduction of these pesticide residues in soils, but there are still high amounts of pesticide residues in soils, which need more remediation processes.     

Alterations of organ histopathology and metallolhionein mRNA expression in silver barb, Puntius gonionotus during subchronic cadmium exposure

Common silver barb,Puntius gonionotus,exposed to the nominal concentration of 0.06 mg/L Cd for 60 d,were assessed for histopathological alterations(gills,liver and kidney),metal accumulation,and metallothionein(MT)mRNA expression.Fish exhibited pathological symptoms such as hypertrophy and hyperplasia of primary and secondary gill lamellae,vacuolization in hepatocytes,and prominent tubular and glomerular damage in the kidney.In addition,kidney accumulated the highest content of cadmium,more than gills and liver.Expression of MT mRNA was increased in both liver and kidney of treated fish.Hepatic MT levels remained high after fish were removed to Cd-free water.In contrast,MT expression in kidney was peaked after 28 d of treatment and drastically dropped when fish were removed to Cd-free water.The high concentrations of Cd in hepatic tissues indicated an accumulation site or permanent damage on this tissue.