石油污染地下水中细菌多样性研究

采集了某废弃炼油厂的石油污染地下水样品,提取水中微生物总DNA,构建细菌16S rDNA克隆文库,并通过16S rDNA序列的系统发育分析,对样品中的细菌种群多样性以及群落结构进行了研究.结果表明,文库中阳性克隆的16S rDNA序列分属10个细菌类群,分别为γ-Proteobacteria(49.1%)、α-Proteobacteria(12.9%)、β-Proteobacteria(11.1%)、Bacteroidetes(9.2%)、Verrucomicrobia(6.7%)、Acidobacteria(2.5%)、δ-Proteobacteria(1.2%)、Actinobacteria(1.2%)、Planctomycetes(0.6%)、Unidentifiedbacteria(5.5%).在这一生态系统中,γ-Proteobacteria类细菌占据主导地位,接近50%,尤其是假单胞菌属(Pseudomonas)微生物在文库中的比例达35.6%.该石油污染地下水样品中细菌与许多其它已知的降解菌亲缘关系较近,如鞘胺醇单胞菌(Sphingomonas)、红球菌(Rhodococcus)和短波单胞菌(B...     

无苯酚培养基分离到的降酚菌多样性的分子分析

用3种不含苯酚的培养基(YPG、10%YPG和LB)从上海焦化厂废水处理系统(A2/O法)好氧池的悬浮污泥分离到24株降酚菌株.通过16SrDNAPCR扩增产物的限制性酶切分析(amplifiedribosomalDNArestrictionanalysis,ARDRA)、ERICPCR指纹图谱分析、多组分苯酚羟化酶大亚基(thelargestsubunitofthemulticomponentphenolhydroxylase,LmPH)基因的PCR扩增及16SrRNA基因测序的方法对这些降酚菌株进行表征.通过ARDRA分型,将这24株降酚菌株分为8个类型;利用ERICPCR可以将这24个菌株分为17种类型,说明同一ARDRA类型内菌株具有多样性.对17个ERIC类型代表菌株的16SrDNA扩增产物进行克隆并测序,测序结果在GenBank和RDP中进行比对.结果表明,与这17个代表菌株同源性最高的菌中,有6株是未见报道具有降酚功能的菌株.在这24株降酚菌中,有19株在苯酚含量为200mg/L的MP培养基中培养5d,苯酚降解率为20%左右,其余5株苯酚降解率达到100%,且均属于Rhodococcus属.本研究在降酚菌生物多样性上作了有益的探索.图2表1参20     

石油降解菌株G5 16S rDNA全序列的系统学分析

用PCR方法扩增了一株石油降解菌株G5的16S rRNA基因全序列，并对其进行了克隆和测序．对该序列在GenBank中的BLAST结果表明，所有与该序列高度同源的序列都是假单胞菌的16S rRNA基因．其中假单胞菌的代表菌株Pseudomonas aeruginosa，P．fluoroscens，P .putida，P.syringae的16S rRNA基因序列与G5的16S rRNA基因序列同源性分别为93．4％，98．4％，96．3％，97．5％．对G5和其他39株假单胞菌的16S rRNA基因序列进行聚类分析，获得的系统发育树与RDP(Ribosomal Database Project)报道的系统发育树基本一致，其中菌株G5与5株P．chlororaphis聚类在一起．图2参7     

贵州茂兰喀斯特森林不同演替下土壤真核微生物多样性

真核微生物是茂兰喀斯特森林土壤微生物中的重要组成部分,在生态系统物质循环和能量流动中扮演着重要角色.为了解茂兰喀斯特森林不同演替阶段下土壤真核微生物群落组成及其多样性,利用18S rDNA高通量测序技术研究了原生林(YSL)、灌木林(GML)、灌木丛(GMC)和草地(CD)这4种演替阶段下土壤真核微生物群落多样性.结果表明,在不同分类水平下,不同演替阶段土壤真核微生物群落组成相似.α多样性指数存在显著差异, Shannon指数与Simpson指数在不同演替阶段表现为:YSLGMCGMLCD. NMDS分析表明不同演替阶段土壤真核微生物群落结构存在差异; LEfSe分析表明,YSL中差异指示种数量高于GML、GML和CD.本研究结果为研究不同演替阶段下土壤真核微生物提供了理论基础.     

新型IEM-UF耦合短程硝化反硝化系统脱氮特性

针对低碳氮比生活污水的特点,提出新型离子交换膜-超滤组合膜（IEM-UF）氮富集短程硝化反硝化脱氮工艺.研究了新型IEM-UF亚硝化反硝化脱氮系统在三阶段运行工况下各反应器的性能及整个系统的脱氮及COD去除效果,同时应用高通量技术探究菌群结构变化对脱氮效果的影响.试验结果表明：C/N为3,亚硝化反应器中DO=0.5mg/L条件下,亚硝化反应器中NO₂^--N积累率仅用19d就达到了90%;在短程反硝化进水流量比为2：1的条件下,COD及NO_x^--N平均去除率分别达到80%和89%以上.TN去除率最高达到64.8%.高通量16S rDNA测序结果表明,三阶段菌群结构变化与系统脱氮效果的变化一致,亚硝化反应3个阶段亚硝化单胞菌Nitrosomonas所占比例分别为3.69%、5.48%和0.53%,反硝化反应3个阶段反硝化菌Dechloromonas、Thauera之和占活性污泥总菌群比例达到33.35%、25.62%、20.52%.     

矿化垃圾生物反应器中的细菌多样性分析

为探究准好氧生物矿化垃圾床处理渗滤液过程中的微生物作用机理,研究建立了16S r DNA克隆文库及PCRRFLP技术以研究矿化垃圾反应器的细菌多样性。结果表明:矿化垃圾反应器细菌具有高度多样性,反应器内有36种细菌分别属于13个纲,变形菌纲占绝对优势占70%(其中β-变形占56.5%,γ-变形菌纲占10%),拟杆菌纲、鞘氨醇菌纲,芽孢杆菌纲也具有一定优势;3%的Nitrosomonas属是渗滤液氨氮转化成亚硝酸盐氮的主要功能微生物,由于亚硝酸盐氧化菌不存在或丰度极低,因此造成反应器内亚硝酸盐积累;Thauera属(17%)和Thiobacillus denitrificans属(10%)是反应器内主要优势微生物属,是反应器内反硝化脱氮的功能微生物,由于Thauera属在好氧条件下具有反硝化特性,Thiobacillus denitrificans为严格自养反硝化菌,因此反应器脱氮主要途径为好氧反硝化、自养反硝化。     

Biodegradation of nicosulfuron by the bacterium Serratia marcescens N80

By enrichment culturing of the sludge collected from the industrial wastewater treatment pond, we isolated a highly efficient nicosulfuron degrading bacterium Serratia marcescens N80. In liquid medium, Serratia marcescens N80 grows using nicosulfuron as the sole nitrogen source, and the optimal temperature, pH values, and inoculation for degradation are 30–35°C, 6.0–7.0, and 3.0% (v/v), respectively. With the initial concentration of 10 mg L^?1, the degradation rate is 93.6% in 96 hours; as the initial concentrations are higher than 10 mg L^?1, the biodegradation rates decrease as the nicosulfuron concentrations increase; when the concentration is 400 mg L^?1, the degradation rate is only 53.1%. Degradation follows the pesticide degradation kinetic equation at concentrations between 5 mg L^?1 and 50 mg L^?1. Identification of the metabolites by the liquid chromatography/mass spectrometry (LC/MS) indicates that the degradation of nicosulfuron is achieved by breaking the sulfonylurea bridge. The strain N80 also degraded some other sulfonylurea herbicides, including ethametsulfuron, tribenuron-methyl, metsulfuron-methyl, chlorimuron-ethyl,and rimsulfuron.     

Characterization and catabolic genes detection of three oil-degrading Rhodococcus spp.北大核心CSCD

Oil pollution is one of the major factors causing environmental deterioration. Bioremediation of oil contaminated environments by microorganisms attracts much research attention. This study aimed to screen efficient oil-degrading bacteria from oil contaminated soil and analyze their characteristics and catabolic genes. Oil-degrading bacteria were screened from oil contaminated soil in minimal medium containing crude oil and identified by morphological, physiological and biochemical characteristics and 16S rDNA sequence analysis. Their growth and degradation characteristics were studied with ultraviolet spectroscopy and GC-MS analysis. The surfactant production was studied by adopting culture method. The major oil-degrading related genes were detected by t he PCR a mplification. As a result, t hree oil-degrading bacteria strains named KB1, 2182 and JC3-47 were isolated from the oil contaminated soil samples. The strains could use crude oil as the sole carbon source to degrade oil with a degrading rate of 41.02%, 32.26% and 55.90%, respectively, when cultured in minimal medium containing crude oil for 3 days. The bacteria were identified to belong to genus Rhodococcus. With 100% similarity of 16S rDNA sequences of the three strains with known ones of Rhodococcus, KB1 was preliminarily identified as Rhodococcus erythropolis, 2182 as Rhodococcus equi, and JC3-47 as Rhodococcus qingshengii. They grew well at 10-50 °C, with the initial pH of 3-9 and the NaCl concentration of 0-5%. The optimal temperature for bacterial growth was 35 °C, 35 °C and 30 °C respectively. KB1 and 2182 could grow at pH 2 and 9.0% of NaCl. The bacteria grew well in broth containing different organic substrates as sole carbon source, such as n-dodecane, n-octadecane, benzene, methylbenzene, xylene and naphthaline. KB1 and JC3-47 could grow well in broth containing pyrene. GC-MS analysis revealed that the bacteria could effectively degrade medium- and long-chain alkane components in crude oil. The bacteria produced biosurfactants and decreased the surface tension of the culture broth. They also showed adhesion activities to n-hexadecane. The oil-degrading related genes such as alkane monooxygenase, aromatic-ring-hydroxylating dioxygenase and catechol dioxygenase genes were detected in all the three strains. Besides, biphenyl dioxygenase genes were detected in KB1 and 2182. The isolated Rhodococcus spp. strains could effectively degrade petroleum hydrocarbons with high adaptabilities to extreme environments such as high salt and low temperature. They are supposed to be applied broadly in the bioremediation of oil contaminated soil in such environments.     

Dynamics of bacterial community of Suillus granulatus mushroom shiro in Pine (Pinus tabuliformis) forests北大核心CSCD

To evaluate bacterial community variation in the mushroom shiro of Suillus granulatus during fruiting, we collected soil samples from the mushroom shiro in the pine (Pinus tabuliformis) forest of mountainous area in Beijing from May to November and evaluated the bacterial community using polymerase chain reaction - denaturing gradient gel electrophoresis (PCR-DGGE). Total soil DNA was extracted using a commercial soil DNA isolation kit. PCR amplification and DGGE were performed using bacterial universal primers 338F and 518R. The specific bands were excised from the gel and sequenced. The results revealed that soil bacterial community maintained considerably high level and changed seasonally with the mushroom fruiting. In total, 53 bands of DGGE profiles were sequenced and divided into 5 phyla (Acidobacteria, Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria and 22 genera (Acidobacterium, Aminobacter, et al). Species from Proteobacteria and Acidobacteria were the dominant bacterial groups sharing considerably high relative abundance, while class a-Proteobacteria was the most abundant group. The variation of the relative abundance of γ-Proteobacteria species was consistent with the mushroom fruiting season. The relative abundance of Acidobacteria species obviously increased before mushroom flush (in July). The fruiting of S. granulatus and the relative abundance of γ-Proteobacteria were correlated with each other. The present study provided a basis for conservation and domestication of mushroom S. granulatus.     

Isolation,identification and desulfurization mechanism of a sulfur-oxidizing bacterium with salt tolerance北大核心CSCD

Micro-organism with efficient desulfurization performance is a key factor in the biological desulfurization technology. This study aimed to seek such a sulfur-oxidizing strain and understand its desulfurization mechanism. Wastewater in a sewage station of natural gas purification plant was used to screen the sulfide-oxidizing strain, and to identify it based on sequence similarity analysis of 16S rDNA and the morphological characteristics. Thiosulfate was used as substrate for investigating the sulfur oxidation performance and salinity tolerance; the OD600, content change of thiosulfate, sulfate, sulfur, pH and total alkalinity in the cultural system were also investigated. The strain DS-B was found to share the highest sequence similarity with Thioalkalivibrio thiocyanoxidans ARh2, therefore determined as Thioalkalivibrio. At the optimum temperature of 35 °C for growth and degradation, the removal efficiency of thiosulfate could reach 98.7% after 7 days. Strain DS-B had strong resistance to thiosulfate, and the optimal concentration of S2O32- was 2 × 104 mg/L. The analysis for sulfur oxides showed that it could oxidize thiosulfate by the pathway of S2O32-→SO42- / S2O32- → S → SO42-. Therefore the strain DS-B is a sulfur-oxidizing bacterium with great application prospect for its strong salt tolerance and conspicuous removal capability for thiosulfate.