Phylogenetic analysis of bacterial community in deep-sea sediment from the western Pacific “warm pool”

A depth profile of bacterial community structure in one deep-sea sediment core of the western Pacific ＂warm pool＂ （WP） was investigated and compared with that in a sediment sample from the eastern Pacific （EP） by phylogenetic analysis of 16S rDNA fragments. Five bacterial 16S rDNA clone libraries were constructed, and 133 clones with different restriction fragment length polymorphism （RFLP） patterns were sequenced. A phylogenetic analysis of these sequences revealed that the bacterial diversity in a sample from the WP was more abundant than that in the EP sample. The bacterial population in the sediment core of WP was composed of eight major lineages of the domain bacteria. Among them the γ-Proteobacteria was the predominant and most diverse group in each section of WP sediment core, followed by the α-Proteobacteria. The genus Colwellia belonging to γ-Proteobacteria was predominant in this sample. The shift of bacterial communities among different sections of the WP sediment core was δ-, ε-Proteobacteria, and Cytopahga-Flexibacteria-Bacteroides （CFB） group. The ratios between them in the bacterial communities all showed inversely proportional to the depth of sediment. The sequences related to sulphate reducing bacteria （SRB） were detected in every section. The bacterial community structure in this sediment core might be related to the environmental characteristics of the surface seawater of the western Pacific WP.     

Phylogenetic diversity of Type I polyketide synthase genes from sediments of Ardley Island in Antarctica

The diversity of modular polyketide synthase (PKS) genes in sediments of Ardley Island in Antarctica, was studied by restriction fragment length polymorphism (RFLP) analysis. Phylogenetic analysis of 14 amino acid (AA) sequences indicates that the identified ketosynthase (KS) domains were clustered with those from diverse bacterial groups, including Cyanobacteria, γ-Proteobacteria, Actinobacteria, Firmicutes, and some unidentified microorganisms from marine sponge, bryozoan and other environmental samples. The obtained KS domains showed 43%-81% similarity at the AA level to reference sequences in GenBank. Six identified KS domains showed diverse sequences of the motif (VQTACSTS) that was used to identify the hybrid PKS/nonribosomal peptide synthetase (NRPS) enzyme complex, and formed a new branch. These results reveal a high diversity and novelty of PKS genes in antarctic sediments.     

宁波北仑港冬季浮游细菌多样性研究

采用分子生物学方法对宁波北仑港冬季水体中的浮游细菌多样性进行了研究.提取水样中细菌基因组总DNA,以细菌16S rDNA通用引物进行PCR扩增,扩增产物经分子克隆、测序与序列分析,对水样中的细菌建立了16S rDNA克隆文库和系统发生树.结果表明,浮游细菌群落具有较高的多样性,34个克隆子分属2个不同的细菌类群,6个克隆子属于未知类群,优势细菌类群为Pro-teobacteria类群（变形菌类群）,占80%;细菌优势类群顺序为β-Proteobacteria类群（32.5%）、γ-Proteobacteria类群（25%）、α-Proteobacteria类群（15%）、ε-proteobacteria类群（7.5%）、Firmicutes类群（厚壁菌类群）（5%）.这一结果与近年来国内外有关港口微生物多样性的报道较为一致.用DNAStar中Clustalw程序从测序的40个克隆子中选出17个OTU进行系统发育分析,同样表明浮游细菌多样性较强.     

辽河口沉积物中古菌和细菌群落结构分析

本研究旨在了解辽河口表层沉积物中古菌和细菌的群落结构组成、多样性及其与环境因子之间的相关性。采用构建古菌和细菌16S rRNA基因文库的方法,应用Illumina Miseq测序技术进行序列分析。结果表明:辽河口表层沉积物中细菌多样性高于古菌多样性,近岸细菌和古菌多样性高于远岸,即河相区（0.8~7.04）细菌多样性高于混合区（13.1~20.7）和海相区（24.2~31.5）;主要古菌群落为奇古菌门（Thaumarchaeota,72.73%）和广古菌门（Euryarchaeota,25.05%）,其中泉古菌门（Crenarchaeota,0.001%）只在河相区站位被发现;细菌群落组成中变形菌门（Proteobacteria,61.94%）为该河口的优势菌群,其次为拟杆菌门（Bacteroidetes,11.21%）和酸杆菌门（Acidobacteria,5.59%）,其他门类如蓝藻门（Cyanobacteria,3.03%）等比例较小。与环境因子的冗余度分析表明:影响表层沉积物中古菌群落分布的主要环境因子依次为氨盐、泥、酸碱度、盐度、电导率和砂,而细菌群落的分布主要受到溶氧、泥、砂、黏土和总磷的影响。由此可见,不同环境条件下微生物的群落结构存在空间异质性,不同微生物对同一环境条件的响应亦不同。     

南海深海氮循环微生物的原位培养与多样性分析

借助自主研发的深海水体原位定植培养系统,在南海3 300 m的深海水体中进行了氮循环微生物的原位培养.通过向富集仓中投加缓释肥(铵盐、硝酸盐和尿素)进行了为期17个月的原位富集.结合高通量测序技术和分离培养的方法,对深海原位富集物及实验室二次富集物进行了微生物多样性分析. 16S rRNA基因高通量测序结果表明,在南海原位富集样品中,细菌以变形菌门丰度最高,富集仓内脱脂棉附着介质和仓内水样中的最优势属分别为希瓦氏菌属(Shewanella)和科韦尔氏菌属(Colwellia);古菌以奇古菌门为主,其中氨氧化古菌(AOA)所占比例很高.通过平板培养分离到17株细菌,主要包括盐单胞菌属(Halomonas)、海杆菌属(Marinobacter)和亚硫酸盐杆菌属(Sulfitobacter)等.通过不同起始氮源(氨氮、硝酸盐、亚硝酸盐)和培养温度(28℃和10℃),对原位富集样品进行实验室二次富集,获得了5个富集菌群.分析发现,它们均具有较好的脱氮效果,可以产生N_2O或N_2;这些菌群均以盐单胞菌属占绝对优势,其次为海源菌属(Idiomarina)或海杆菌属,其中盐单胞菌和海源菌成功获得分离.单菌功能验证表明,分离获得的盐单胞菌具有好氧反硝化能力,其在原位环境下反硝化活性尚有待确认.     

北极海洋沉积物细菌和古菌群落结构分析

This study was to investigate bacterial and archaeal community structure of pan-Arctic Ocean sediments by pyrosequencing.In total,investigation of three marine sediments revealed 15 002 bacterial and 4 362 archaeal operational taxonomic units(OTUs) at the 97% similarity level.Analysis of community structure indicated that these three samples had high bacterial and archaeal diversity.The most relatively abundant bacterial group in Samples CC1 and R05 was Proteobacteria,while Firmicutes was dominant in Sample BL03.Thaumarchaeota was the most relatively abundant archaeal phylum in Samples CC1 and R05,and the relative abundance of Thaumarchaeota was almost as high as that of Euryarchaeota in Sample BL03.These two phyla accounted for nearly 100% of the archaeal OTUs.δ-Proteobacteria and γ-Proteobacteria were the two most relatively abundant classes at Proteobacterial class level,and their relative abundance was more than 60% in Samples CC1 and R05.There were also differences in the top 10 relatively abundant bacterial and archaeal OTUs among the three samples at the 97% similarity,and only 12 core bacterial OTUs were detected.Overall,this study indicated that there were distinct microbial communities and many unique OTUs in these three samples.     

海南红树林根系土壤中可培养细菌的多样性分析

为研究海南红树林根系土壤细菌多样性,充分发掘中国红树林特有微生物资源,作者于2012年12月份从10份采集于海南北港岛红树林根系淤泥样品中分离可培养海洋细菌,并基于16S rRNA基因序列分析的方法对样品中的细菌多样性进行研究。共分离得到122株海洋细菌,经并菌后选取其中的70株代表性菌株测序,结果发现它们分为4个类群:变形菌门(主要为γ-变形菌纲)(Proteobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)和放线菌门(Actinobacteria),分别占66%,19%,11%和4%。共代表30个属47个种,其中有9个潜在新种和2个潜在新属。本研究结果表明,北港岛红树林中存在着丰富的细菌多样性,很多类群可能是红树林特有细菌物种。     

厦门近岸水域海洋附着细菌的群落结构及其动态变化

为研究海洋附着细菌的群落结构及动态变化,在厦门近岸海区进行挂板实验.将无菌玻璃板浸没于海水中,连续放置14 d.分别于放置1 h和7、14 d后取玻璃板上附着生物样品.用细菌通用引物构建16S rRNA基因克隆文库,每个克隆文库随机挑选约40个克隆子测序,序列同源性分析和系统进化分析结果表明,所有的克隆子可分为六大类群:γ-变形菌纲(γ-Proteobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、α-变形菌纲(α-Proteobacteria)、蓝细菌门(Cyanobacteria)和真核硅藻类叶绿体,各类群分别占42.0%、4.5%、2.2%、2.2%、1.1%和45.0%.γ-变形菌纲变形斑沙雷氏菌(Serratia proteamaculans)为优势附着细菌,占测序克隆子的31.5%.这类细菌在1 h样品中的比例超过一半,说明变形斑沙雷氏菌在生物膜形成初期发挥着重要作用.随着挂板时间延长,检测到的细菌类群有所增加:附着7 d后检测到拟杆菌门细菌,附着14 d后检测到厚壁菌门细菌.γ-变形菌纲细菌所占比例随挂板时间的延长而逐渐降低,从挂板1 h的81%降至7 d的21%,14 d的18%.另外,在各阶段的附着样品中,都检测到较多的真核克隆子序列,约占16%～64%.本研究为进一步阐明海洋附着细菌的附着动态及其在生物膜形成过程中的作用奠定了基础.     

北黄海微表层细菌丰度与可培养细菌群落结构分析

为分析海洋微表层这一特殊生境中的细菌类群,于2010—2011年4个航次对北黄海微表层和次表层海水中的总菌丰度、可培养细菌丰度和群落结构进行了分析。采用流式细胞仪测定总菌丰度,平板计数法测定可培养细菌丰度,PCR-16SrDNA分析可培养细菌的群落结构。结果表明,调查海域微表层海水总菌均值为1.76×106 cell/mL,次表层海水总菌均值为1.07×106 cell/mL。可培养细菌丰度范围是1.00×102~1.70×106 CFU/mL,微表层和次表层可培养细菌所占总菌数量的百分比分别为13.05%和0.45%。微表层对总菌的富集因子(EF)均值为2.02,可培养细菌的EF均值为74.16。PCR-16SrDNA序列分析结果表明,该海域可培养细菌分属变形菌门(Proteobacteria)(94.34%)、厚壁菌门(Firmicutes)(1.89%)、拟杆菌门(Bacteroidetes)(1.89%)和放线菌门(Actinobacteria)(1.89%)4个类群。本研究初步发现,微表层对细菌具有较强的聚集作用,尤其对可培养细菌聚集作用更为明显。微表层中可培养细菌的群落结构与次表层有所不同,其种类丰富,来源和功能多样。由此可见,微表层独特的生境成就了其独特的微生物类群,其生态功能有待于进一步研究。     

Microbial community respiration and structure of dead zone sediments of Omura Bay, Japan

Monitoring sediment microbial community metabolism and structure is instrumental to understanding biogeochemical processes in and ecological impacts on bottom environments. The aim of this study was to determine potential community respiration and to reveal community dynamics of the microorganisms in the dead zone sediments of Omura Bay, Japan. The bay is highly enclosed and develops severe hypoxia in the central regions every summer. We collected sediment core samples from the center of the bay during hypoxia, estimated sediment oxygen consumption by using an adapted in vivo electron transport system activity (in vivo ETSA) assay, enumerated abundance of bacteria, and analyzed bacterial community structure by automated ribosomal intergenic spacer analysis. Higher ETSA and bacterial diversity were found in upper sediments (within 3?cm depth) from the center than the fringe of the bay. Sediment bacterial community structure of the bay center was distinct from that of the fringe. From these results, upper sediment in the dead zone of Omura Bay was characterized by (1) greater community respiration and (2) greater diversity of bacterial components compared with the non-hypoxic sediment of the bay fringe. These characteristics have important implications for understanding the interaction between microbial communities and the development of hypoxia in Omura Bay.     

Microbial diversity in polluted harbor sediments I: Bacterial community assessment based on four clone libraries of 16S rDNA

Bacteria, as the most abundant sediment organism, play a major role in the fate of pollutants. Therefore, many pollutant-related bacteria have been studied in harbor sediments, yet the entire bacterial profiles have not been reported. The bacterial diversity and community structures from sediments in Victoria Harbor (Hong Kong), including two polluted (VH and VHW) and two adjacent (open oceanic, TLC; estuary discharge affected, PC) sites, were characterized by analyses of four 16S rDNA clone libraries. Upon comparisons of RFLP patterns from 254 clones in the libraries, 178 unique phylotypes were retrieved. LIBSHUFF and Rarefaction analyses indicated that the sediment bacterial communities at the four sites showed high 16S rDNA richness and were significantly different from each other. Phylogenetic analysis of full-length 16S rDNA revealed 19 bacterial phyla in Victoria Harbor sediments. γ- and δ-proteobacteria, holophaga/acidobacteria, and planctomycetales were recorded in all the libraries. In addition, γ- and δ-proteobacteria were dominant at all sites (33.33–11.67%). Besides these two phyla, ε-proteobacteria, firmicutes, aminobacterium, holophaga/acidobacteria and bacteroidetes were judged to be major components of a given library since they constituted 10% or more of the total OTUs of the given library. The cyanobacteria, verrucomicrobia, β-proteobacteria, aminobacterium, chlorofiexi, and candidate division OP1, OP8 were detected in minor proportions in various libraries. A portion of the clones were only distantly related to sequences in the GenBank, suggesting bacteria in Victoria Harbor sediments were unique and diversified.     

人工饲料饲养的对虾肠道菌群和水体细菌区系的研究

对人工饲料饲养的凡纳滨对虾Litopenaeus vannamei成虾肠道菌群组成及其水体细菌区系组成进行了研究.分离纯化后的革兰氏阴性菌鉴定到种的水平.对虾肠道菌群主要由弧菌Vibrio spp.、希瓦氏菌属Shewnella spp.、嗜氢菌属Hydrogenophaga spp.、伯克霍尔德氏菌Burkholderia spp.、气单胞菌Aeromonas spp.、食酸菌Acidovorax spp.和芽孢杆菌Bacillus spp.组成.水体细菌区系主要包括弧菌Vibrio spp.、希瓦氏菌属Shewnella spp.、艾肯菌Empedobacter spp.、无色杆菌Achromobacter spp.和芽孢杆菌Bacillus spp..肠道菌群和水体细菌区系的优势菌是副溶血弧菌Vibrio parahaemolyticus和芽孢杆菌Bacillus spp..利用Shannon-Wiener指数进行多样性分析,发现虾肠道多样性指数（H）和均匀度（J）高于水体细菌区系,而丰度（D）较低.     

Characterization of bacterial communities associated with seasonal water masses from Tongyoung in South Sea of Korea

Understanding the patterns of microbial diversity and their composition in seawater is necessary to assess the impacts of microbes on marine ecosystem. Although the potential roles of microbes in the ocean have been studied, their diversity and relationship with environmental factors remain unclear. In this study, we adopted a 16S rRNA gene tag-pyrosequencing technique to investigate the bacterial communities associated with two different water masses from Tongyoung in the South Sea of Korea. Our results revealed highly diverse bacterial communities up to 900 Operational Taxonomic Units (OTUs) estimated from each seawater which was collected in the month of March and May, when the environmental conditions including temperature differed significantly: 7.2°C and 17.6°C in March and May, respectively. Altogether, 13 bacterial phyla were recovered from the seawater, of which Proteobacteria was the most dominant group. In addition, the value of the Shannon index, which measures the evenness of the distribution of individuals among OTUs, in May is higher than that in March, indicating that it displays a wider diversity of bacteria. Interestingly, the proportion of pathogenic bacteria was significantly increased in the month of May compared to March, suggesting that pathogenic bacteria were increasingly emerging in May. In particular, Pseudoalteromonas and Vibrio spp. were determined as major pathogenic bacteria from both water masses, of which Vibrio spp. were dominant.     

西太平洋深海海水中乙醛降解菌多样性及群落结构研究

As a major aldehyde pollutant widely existing in industry and our daily life, acetaldehyde is more and more harmful to human health. As characteristic habitat niche, bacteria from deep sea environments are abundant and distinctive in heredity, physiology and ecological functions. Thus, the development of acetaldehyde-degrading bacteria from deep sea provides a new method to harness acetaldehyde pollutant. Firstly, in this study,acetaldehyde-degrading bacteria in the deep sea water of the West Pacific Ocean were enriched in situ and in the laboratory respectively, and then the diversity of uncultured bacteria was studied by using 16 S r RNA genes. Then acetaldehyde-degrading strains were isolated from two samples, including enrichment in situ and enrichment in laboratory samples of deep sea water from the West Pacific Ocean using acetaldehyde as the sole carbon source,and then the ability of acetaldehyde degradation was detected. Our results showed that the main uncultured bacteria of two samples with different enrichment approaches were similar, including Proteobacteria,Actinobacteria, Firmicutes, Cyanobacteria, but the structure of bacterial community were significant different.Four subgroups, α, γ, δ and ε, were found in Proteobacteria group. The γ-Proteobacteria was dominant(63.5%clones in laboratory enriched sample, 75% clones in situ enriched sample). The species belonged to γ-Proteobacteria and their proportion was nearly identical between the two enrichment samples, and Vibrio was the predominant genus(45% in laboratory enriched sample, 48.5% in situ enriched sample), followed by Halomonas(9% in situ enriched sample) and Streptococcus(6% in laboratory enriched sample). A total of 12 acetaldehyde-degrading strains were isolated from the two samples, which belonged to Vibrio, Halomonas,Pseudoalteromonas, Pseudomonas and Bacillus of γ-Proteobacteria. Strains ACH-L-5, ACH-L-8 and ACH-S-12,belonging to Vibrio and Halomonas, have strong ability of acetaldehyde degradation, which could tolerate 1.5 g/L acetaldehyde and degrade 350 mg/L acetaldehyde within 24 hours. Our results indicated that bacteria of γ-Proteobacteria may play an important role in carbon cycle of deep sea environments, especial the bacteria belonging to Vibrio and Halomonas and these strains was suggested for their potentials in government of aldehyde pollutants.     

天然海水中微生物膜对碳钢腐蚀行为的影响

通过对比碳钢在天然海水和灭菌海水中的腐蚀行为,研究海洋微生物对碳钢材料的腐蚀行为的影响.结果表明,海洋微生物可在碳钢表面形成微生物膜,其对碳钢腐蚀速率的影响及作用机理与微生物的种类密切相关.在腐蚀初期,海洋微生物膜对碳钢的腐蚀起到了抑制作用,主要由多种好氧微生物协同作用的结果;在腐蚀后期,随着海洋微生物膜增厚导致厌氧腐蚀的出现从而加速腐蚀,主要是硫酸盐还原菌等厌氧微生物的作用.     

The bacterial diversity in surface sediment from the South China Sea

16S rDNA sequencing results from this study and literatures demonstrate that sediment bacteria in the South China Sea (SCS) were very diverse,which contained 22 of the 24 phyla of bacteria investigated from marine sediment,however,it was very imbalance among stations.So bacterial diversity from 15 samples which covered a wide range of sediment types from 20 to 3 888 m in depth was studied in DGGE (denature gradient gel electrophoresis) in this paper.The DGGE results indicate that both sediment bacterial diversity and diversity difference among stations were significant.Thirty representative and differential fingerprints among samples were recovered and sequenced,phylogenetic analysis indicates that they may belong to Proteobacteria (α-,β-,γ-,δ-,ε-),Planctomycetes,Firmicutes,Chloroflexi,Acidobacteria,Actinobacteria,Nitrospirae,Gemmatimonadetes,candidate division WS3 and so on,of which,Gemmatimonadetes and candidate division WS3 bacteria were first detected in SCS sediment.This study also shows that bacterial diversity analysis based on DGGE was more potential than traditional 16S rDNA clone library in multiple sample analysis.     

乳山湾外海溶氧低值区细菌多样性初步研究

采用化学裂解法从乳山湾外海溶氧低值区不同溶解氧质量浓度(3.7～7.0mg.L-1)的6个站位海水样品中提取了环境DNA样品。以试剂盒纯化后的DNA样品为模板扩增其16SrRNA基因V3区,通过变性梯度凝胶电泳、分子文库构建及DNA测序对溶氧低值区海水中的细菌群落结构进行研究。结果表明,乳山湾外海溶氧低值区不同溶解氧质量浓度的6个站位底层海水样品中的细菌群落结构是相似的,它们均由隶属于Alteromonas(交替单胞菌属)、Salegentibacter(需盐杆菌属)等10个属的18种细菌组成。系统发育分析发现这些细菌分别属于α变形菌纲(2种)、γ变形菌纲(12种)和黄杆菌纲(3种)三个大类。在乳山湾外海溶氧低值区的海水样品中细菌多样性最高的类群是γ变形菌纲。     

海洋源拮抗细菌的分离、筛选及1株拮抗芽孢杆菌的鉴定

对来自于不同海域的红树树皮、海苔(海洋生物样)和海泥(海洋底质)样品进行细菌分离,并采用管碟法测试了分离出的海洋细菌对金黄色葡萄球菌Staphylococcus aureus、枯草芽孢杆菌Bacillus subtilis、大肠杆菌Escherichia coli、白色念珠菌Candida albicans、黑曲霉As...     

东太平洋中国多金属结核区锰结核样品中微生物群落结构的研究

通过非培养手段研究了东太平洋中国多金属结核区ES0303站点锰结核样品中的微生物群落结构.细菌16S rRNA基因克隆文库的研究结果表明：结核内细菌种群结构复杂,微生物种类丰富且各种群丰度不一（61个OTUs）,其中变形杆菌类群为优势种群,占所有细菌克隆子比例的64%,且主要分布于β/γ-、α-和δ-等3类变形杆菌（Proteobacteria）亚群之中,占比分别为34%、18%和12%.此外还存在包括酸杆菌（Acidobacteria）,放线菌纲（Actinobacteria）,绿弯菌门（Chloroflexi）,厚壁菌门（Firmicutes）,浮霉菌门（Planctomycetes）等在内的细菌类群的分布,克隆子比例依次为9%、7%、8%、2%和5%.古菌16S rRNA基因克隆文库的研究结果表明：古菌的群落结构单一（仅12个OTUs）,全部是由泉古菌海洋类群I（crenarchaeote marine group I,MGI）组成;其中MGI-η类群最为丰富,达到44%,而MGI-α、MGI-ζ和MGI-ε类群的克隆子比例分别为25%、18%和9%,另外还发现2个新的MGI分类类群.相关克隆子的数据库比对和系统发育树分析表明,并未发现已报道的直接参与铁锰氧化还原相关类群的存在,但它们大多数与来自多金属结核来源或深海来源的不可培养微生物具有较高的同源性.进一步的分析表明,锰结核内存在相当数量的氨氧化菌、硫酸盐还原菌、酸杆菌等能够改变pH值的细菌和古菌类群的存在,意味着它们可能在锰结核的形成过程中起到重要的作用.     

Interactions of bacteria with diatoms: influence on natural marine biofilms

Interactions between microfouling components influence the biofilm community and the cascading events, thus playing an important role in the biofouling process. Bacteria and diatoms are among the dominant forms reported in biofilms. Experiments were carried out with natural marine biofilms from a tropical monsoon‐influenced environment to evaluate the interactions between bacteria and diatoms through application of antibiotics (streptomycin and chloramphenicol). Overall, chloramphenicol inhibited diatom communities, whereas streptomycin did not. These antibiotic‐mediated changes in the fouling diatom community were consistent across the seasons. However, the rates at which the fouling communities changed depended on the initial species composition. It was also observed that elevated nutrient concentrations overrode the inhibitory effect of chloramphenicol. Maximum Vibrio enhancement was observed in the enriched conditions during the pre‐monsoon and unenriched conditions during the monsoon (with naturally elevated background nutrient concentrations), highlighting the relevance of nutrient concentrations for Vibrio spp. This has interesting implications for antibiotic‐mediated interactions between fouling diatom and bacterial communities under differing nutrient regimes. Although this study indicates the relevance of ‘cross‐talk’ at the microfouling level, understanding the effects of additional microbial products (e.g. bacteriocins and peptidoglycan) on these community‐level interactions will represent a step forward.