北极海洋沉积物中锰细菌的分离与系统发育

对中国第二次北极科学考察采集的北极海洋沉积物中的锰细菌进行了筛选、分离和系统发育分析。根据其在筛选平板上菌落的形态学特征,分别从站位P11和S11采集的沉积物中分离到了21株和19株锰细菌。系统发育分析表明,两个站位的锰细菌群落组成有着明显的差别。站位P11分离的可培养锰细菌主要由细菌域（Bacteria）中变形杆菌门的γ-变形杆菌纲（γ-Proteobacteria）和放线菌纲（Actinobacteria）组成,二者分别占86％和14％;γ-变形杆菌纲主要包括嗜冷杆菌属（Psychrobacter）、希瓦氏菌属（Shewanella）、假交替单胞菌属（Pseudoaheromonas）、不动杆菌属（Acinetobacter）、海杆菌属（Marinobacter）,其中以嗜冷杆菌属为主,其比例可达67％。从站位S11分离到的可培养锰细菌主要包括细菌域中变形杆菌门的α-变形杆菌纲（α-Proteobacteria）和γ-变形杆菌纲以及拟杆菌门（Bacteroides）中的黄杆菌纲（Flavobaeteria）;γ-变形杆菌纲主要包括希瓦氏菌属、海单胞菌属（Marinomonas）和交替单胞菌属（Aheromonas）,α-变形杆菌纲主要为鞘氨醇单胞菌属（Sphingomonas）。实验菌株均对Mn^2＋有着较强的抗性,其中以菌株Marinomonas sp．S11-S-4耐受性最高。     

第六次北极科学考察海洋沉积物可培养细菌的多样性分析

【目的】研究北极海洋沉积物可培养细菌的菌种资源多样性。【方法】采用海水Zobell2216E培养基和涂布平板法对第六次北极科学考察获得的海洋沉积物开展细菌分离培养,通过16S rRNA基因系统发育分析了解可培养细菌的多样性。【结果】根据菌落形态特征,从40个站位的北极海洋沉积物样品中共分离并获得16S rRNA基因有效序列的细菌达445株;基于16S rRNA基因的相似性分析与系统发育研究结果表明,分离获得的细菌分属于细菌域的4个门、6个纲、13个目、28个科、49个属、91个种,其中γ-Proteobacteria占大多数;有12株与模式菌株的16S rRNA基因序列相似性小于97%,可能代表了6个潜在的细菌新物种;此次获得的细菌种类组成与以往第五次北极科考获得的相比,在属水平上差异较大。【结论】北极海洋沉积物中存在着丰富的微生物菌种资源,具有很多新型微生物仍未被发现,是亟待开发的微生物资源宝库。     

东北太平洋深海沉积物细菌多样性

采用两种方法提取中国结核合同区东区沉积物不同层次总DNA,通过克隆测序构建了含有79个克隆子的细菌16S rRNA基因文库,分析了该海域沉积物中细菌的多样性.79个克隆在系统发育树中形成了11个大分支,包括Gamma proteobacteria(22.8%),Alpha proteobacteria(16.5%),Planctomycetacia(7.6%),Delta proteobacteria(6.3%), Nitrospira(6.3%),Actinobacteria(6.3%),Beta proteobacteria(5%),Acidobacteria(5.1%),Sphingobacteria(3.8%),Firmicutes(2.5%),Other bacteria(17.7%),其中Gamma proteobacteria在总文库中所占比例最高,该分支细菌在0～2cm、4～6cm层也是优势菌种.Gamma proteobacteria中假单胞菌(Pseudomonas)为优势属(22.2%).各个层次中所含细菌类群有所不同,Alpha proteobacteria、Gamma proteobacteria、Delta proteobacteria 、Planctomycetacia、Nitrospira 、Actinobacteria和Acidobacteria为三层样品共有类群.     

黄海海域海洋沉积物细菌多样性分析

【背景】海洋独特的环境造就了海洋生物的多样性,海洋沉积物中细菌对海洋环境具有至关重要的作用。【目的】研究陆地土壤和海洋沉积物间细菌群落相似性和差异性,以便更好地认识海洋细菌多样性,深入了解沉积物细菌在海洋环境中的潜在作用。【方法】从中国黄海海域及大连市大黑山脚下分别采集样品,以陆地土壤为对照,采用16SrRNA基因高通量测序技术分析海洋沉积物的细菌群落结构。【结果】海洋沉积物样品中芽孢杆菌纲(Bacilli)、鞘氨醇单胞菌属(Sphingomonas)和芽孢杆菌属(Bacillus)丰度高于陆地土壤样品;海洋沉积物中亚硝化单胞菌(unculturedbacterium f. Nitrosomonadaceae)和厌氧绳菌(uncultured bacterium f. Anaerolineaceae)丰度虽低于陆地土壤,但丰度值也均高于1%;样品分类学统计显示酸杆菌门(Acidobacteria)在海洋沉积物和陆地土壤样品中的序列丰度比例都较大,鞘氨醇单胞菌属(Sphingomonas)在海洋沉积物样品中的序列丰度大于陆地土壤样品。【结论】海洋沉积物细菌多样性可作为海洋环境恢复情况的重...     

细菌系统发育

系统发育（Phylogeny）指生物种族的进化历史［1］。19世纪末到20世纪初,微生物学家与其它生物学家一样承认进化,认定检测细菌之间的进化关系非常重要．从Beijerinck、kluyver至VanNiel,在研究细菌的自然关系方面尽了很大努力,但其结果是光依靠表型特征达不到自然分类的目的．VanMel和他以前的学生StainierRY不得不放弃这个想法,并认为“微生物进化的主要途径可能是永远不得而知的”［2］。后来有人提出原核生物与真核生物两分叉的看法。1969年W：lllttakerPd’］提出生物的五界分类系统,即原核界、原生界、植物界、真菌界和动物…     

北极高维度海域海冰嗜冷菌系统发育多样性及其低温水解酶分析

应用样品直接稀释涂布平板、-1℃富集培养和-20℃冷冻24h后富集培养等3种方法,从北极加拿大海盆和格陵兰海的高纬度海域(77°30′N~81°12′N)海冰中分离到37株嗜冷菌。根据其16S rDNA全长序列所进行的系统发育分析表明,分离菌株分属于γ_变形细菌群(γ_Proteobacteria)的Colwellia、Marinobacter、Shewanella、Thalassomonas、Glaciecola、Marinomonas、Pseudoalteromonas和嗜纤维菌_曲挠杆菌_拟杆菌群(Cytophaga_Flexibacter_Bacteroide,CFB)的Flavobacterium、Psychroflexus等9个属。其中有9株菌的16S rDNA序列与已明确鉴定种的相似性在93.4%~96.9%,为潜在的新种。北极加拿大海盆海冰细菌BSi20002与南极威德尔海海冰细菌Marinobactersp.ANT8277的16S rDNA序列相似性为100%,表明在种水平上南、北两极也存在相同的细菌。分离的嗜冷菌在4℃条件下能产生多种大分子物质水解酶类,其中62.6%、51.4%和40.5%的菌株分别能水解Tween_80、明胶和淀粉。     

乌梁素海富营养化湖区浮游细菌多样性及系统发育分析

水生生态系统富营养化与细菌群落之间的关系尚不明确。本文通过构建和分析16S rRNA基因片段克隆文库, 以期揭示乌梁素海富营养化水体细菌的多样性及其系统发育关系, 并探讨富营养化与细菌多样性之间的关系。利用Hae III对文库中的87个克隆子进行单酶切, 产生了23种带型, 文库覆盖度达到了73.6%, 反映出文库有较好的代表性。选择每种OTU的一个代表克隆进行测序分析, 基因序列系统发育分析结果表明, 乌梁素海中多数细菌与淡水生态系统中常见的细菌门类相同, 即α-, β-, γ-Proteobacteria, Bacteroidetes, Actinobacteria, 它们分别占总菌数的10.3%、41.4%、4.6%和6.9%, 其中β-Proteobacteria和Bacteroidetes是优势细菌类群。与典型淡水生态系统细菌群落组成不同的是, 乌梁素海中存在约10.3%的轻度嗜盐碱细菌。水体中83.9%的细菌与已培养的细菌的同源性低于97%, 其中58.9%的细菌未能鉴定到属; 其余总菌数16.1%的克隆与具有降解污染物生物活性的已知菌相近。Bacteroidetes、Firmicutes和β-Proteobacteria中的某些类群成为优势菌群可能是对乌梁素海水体富营养化的响应。     

新疆沙湾冷泉沉积物的细菌系统发育多样性

为了解新疆沙湾冷泉沉积物的细菌群落组成与类群多样性,利用免培养方法直接从沙湾冷泉沉积物中提取环境总DNA,构建细菌16S rRNA基因文库。对随机挑选的241个细菌阳性克隆子进行HaeIII酶切分型得到86个可操作分类单元(OTUs),系统发育分析将其归为11个门:放线菌门(Actinobacteria),酸杆菌门(Acidobacteria),拟杆菌门(Bacteroidetes),绿菌门(Chlorobi),蓝细菌门(Cyanobacteria),厚壁菌门(Firmicutes),芽单胞菌门(Gemmatimonadetes),硝化螺旋菌门(Nitrospirae),变形菌门(Proteobacteria),浮霉菌门(Planctomycetes),疣微菌门(Verrucomicrobia)。其中酸杆菌门和变型菌门为优势类群,分别占细菌克隆文库的48%和25%。超过1/3的OTUs序列与GenBank中已存序列具有较低相似性(相似性小于95%)。此外20%左右的克隆子与固氮细菌和硝酸盐氧化细菌相关。研究结果表明,新疆沙湾冷泉沉积物中细菌种类丰富,代谢类型多样而且存在大量未知类群。     

油樟内生芽孢细菌的系统发育多样性

摘要：【目的】为了解油樟产芽孢内生细菌的多样性。【方法】采用改良的牛肉膏琼脂培养基分离、去除冗余及芽孢染色,测定所得产芽孢内生细菌的16S rRNA基因,进行系统发育分析。【结果】40株产芽孢内生细菌数量占分离得到的内生细菌总数的38.1％,其中根、茎、叶中分别分离得到24株、7株和9株。16S rRNA基因序列系统发育分析结果表明,35株菌可能分属于Bacillus、Lysinibacillus、Paenibacillus属的16个种,还有5株菌的序列与数据库中典型菌株序列相似性低于97％,代表着潜在新类群的存在。【结论】从油樟3个部位分离出的产芽孢内生细菌存在明显的系统发育多样性,而且3个部位分离出的产芽孢内生细菌区系既呈现出一定程度的细菌区系相似性,又表现出器官细菌区系的特异性。     

湛江硇洲岛海葵相关可培养细菌系统发育多样性

【目的】了解湛江硇洲岛海葵样品中相关可培养细菌的多样性。【方法】运用纯培养法和基于16S rRNA基因序列的系统发育分析对样品中可培养细菌多样性进行研究。【结果】用补充0～2.0 mol/L NaCl 的MA、ISP 2、NA、SWA和HAA培养基从海葵样品中分离到126株细菌,通过形态观察和部分生理生化实验去冗余,选取42株具有代表性的菌株进行基于16S rRNA基因序列的系统发育多样性分析。结果表明,42个分离菌株可分为25个物种,属于3个大的系统发育类群(Firmicutes,Gamma-Proteobacteria,Actinobacteria)、11个科、18个属。多数菌株属于Firmicutes门（17株,40.5％）和Gamma-Proteobacteria亚门（14株,33.3％）。这些分离菌株中,至少有6个菌株可能代表6个不同属的6个新物种：JSM 071004、JSM 071068、JSM 071073、JSM 072002、JSM 073008和JSM073097分别代表Bacillus、Halobacillus、Piscibacillus、Pontibacillus、Alteromonas和Nocardiopsis属的新物种。【结论】从以上结果可以看出,湛江硇洲岛海葵中存在较为丰富的微生物物种多样性和系统发育多样性,并且潜藏着较多的新的微生物类群（物种）。     

Bacterial diversity in surface sediments from the Pacific Arctic Ocean

In order to assess bacterial diversity within four surface sediment samples (0–5 cm) collected from the Pacific Arctic Ocean, 16S ribosomal DNA clone library analysis was performed. Near full length 16S rDNA sequences were obtained for 463 clones from four libraries and 13 distinct major lineages of Bacteria were identified (α, β, γ, δ and ε-Proteobacteria, Acidobacteria, Bacteroidetes, Chloroflexi, Actinobacteria, Firmicutes, Planctomycetes, Spirochetes, and Verrucomicrobia). α, γ, and δ-Proteobacteria, Acidobacteria, Bacteroidetes, Actinobacteria were common phylogenetic groups from all the sediments. The γ-Proteobacteria were the dominant bacterial lineage, representing near or over 50% of the clones. Over 35% of γ-Proteobacteria clones of four clone library were closely related to cultured bacterial isolates with similarity values ranging from 94 to 100%. The community composition was different among sampling sites, which potentially was related to geochemical differences.     

Phylogenetic analysis of Archaea in the deep-sea sediments of west Pacific Warm Pool

Archaea are known to play important roles in carbon cycling in marine sediments. The main compositions of archaeal community in five deep-sea sediment samples collected from west Pacific Warm Pool area (WP-0, WP-1, WP-2, WP-3, WP-4), and in five sediment layers (1 cm-, 3 cm-, 6 cm-, 10 cm-, 12 cm- layer) of the 12 cm sediment core of WP-0 were checked and compared by denaturing gradient gel electrophoresis and 16 S rRNA gene sequencing. It was revealed that all the deep-sea sediment samples checked contained members of non-thermophilic marine group I crenarchaeota as the predominant archaeal group. To further detect groups of archaea possibly relating with C1 metabolism, PCR amplification was carried out using primers targeting methane-oxidizing archaea. Although no methane-oxidizing archaea was detected, a group of novel archaea (named as WPA) was instead identified from all these five WP samples by clone analysis. They could be placed in the euryarchaeota kingdom, separated into two distinct groups, the main group was peripherally related with methanogens, the other group related with Thermoplasma. The vertical distributions of WPA, archaea and bacteria along the WP-0 sediment column were determined by quantitative-PCR. It was found that bacteria dominated at all depths, the numbers of bacteria were 10–10⁴ times more than those of archaea. The proportion of archaea versus bacteria had a depth related increasing tendency, it was lowest at the first layer (0.01%), reached highest at the 12 cm- layer (10%). WPA only constituted a small proportion of the archaeal community (0.05% to 5%) of west Pacific Warm Pool sediment.Peng Wang and Xiang Xiao contributed equally to this paper     

Measurement of bacterial production in stream-bed sediments via leucine incorporation

Abstract: The leucine incorporation technique was evaluated and optimised for measuring bacterial production in stream-bed sediments. The original procedure was modified in order to obtain reliable production estimates in this habitat. This included the use of higher leucine concentrations (50 μmol l⁻¹) for obtaining substrate saturation, and an enhanced protein extraction procedure after sample fixation. The leucine method was combined with a perfused core technique. Water containing ¹⁴C-labelled leucine was perfused up through sediment cores, enabling the measurement of bacterial production in an experimental situation resembling natural conditions in the stream bed. Bacterial production in the Breitenbach, a small upland stream in Central Germany, showed a high degree of spatial variability in the sandy stream bed. It was related closely to sediment organic matter content, whereas varying perfusion rates had less influence. Annual bacterial production was estimated at about 200 g C m⁻², demonstrating the potential for bacteria to act as a food resource for benthic fauna in this stream.     

Denaturing gradient gel electrophoresis (DGGE) analysis of bacterial community composition in deep-sea sediments of the South China Sea

The South China Sea, which is one of the largest marginal seas in the world, is predicted to have suitable accumulation conditions and exporting prospects for natural gas hydrate. The aim of this study was to explore the bacterial community composition of deep-sea sediments from such an ecosystem. DNA was extracted by five different methods and used as templates for PCR amplification of the V3 regions of the 16S rRNA gene. Denaturing gradient gel electrophoresis (DGGE) was used to separate the amplified products and analyse the 16S rRNA gene diversity of sediment samples. The results of DGGE indicated that the bacterial community composition is influenced by DNA extraction methods. Sequencing dominant bands demonstrated that the major phylogenetic groups identified by DGGE belong to Proteobacteria, Bacteroidetes, gram-positive bacteria and Archaea. Integrating different DNA extraction procedures are needed to analyse the actual bacterial diversity from environment when the amplification of 16S rRNA gene and construction of representative clone library were adopted.     

典型高原湖滨带底泥细菌群落结构及多样性特征

【背景】高原湖泊的富营养化日趋严重,而湖滨带作为湖泊的保护屏障对外源污染物具有拦截净化等作用,水环境变化则会对底泥细菌产生深刻影响。【目的】探究高原湖滨带底泥细菌群落结构特征及与水体富营养化之间的联系。【方法】基于16S rRNA基因高通量测序技术分析了阳宗海南岸湖滨带8个不同样点的底泥细菌群落结构及多样性,并结合样品水体环境因子,采用主成分分析(PCA)和冗余分析(redundancy analysis,RDA)探讨了水体富营养化对底泥细菌群落结构及丰富度的影响。【结果】湖滨带底泥细菌与水体富营养化程度存在响应关系,在水体富营养化程度高的区域(S3)细菌丰富度较高,操作分类单元(operationaltaxonomicunits,OTU)高达1473。反之,在富营养化程度低的区域(S1)细菌丰富度较低,OTU为730。阳宗海南岸湖滨带底泥中主要优势菌门为变形菌门(Proteobacteria)和绿弯菌门(Chloroflexi),含有少量的放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)和厚壁菌门(Firmicutes);绿弯菌门(Chloroflex...     

Phylogenetic diversity of bacteria in the Arctic Ocean sediments neighboring the Bering Strait

To expand investigations and insights into the phylogenetic diversity of bacteria inhibiting seafloor biosphere, six Arctic Ocean sediments neighboring the Bering Strait were sampled and their bacterial diversities were investigated by pyrosequencing of 16S rRNA genes. A total of 157,454 trimed sequences were obtained, resulting in 9413 OTUs at the 97% sequence identity (OTU_3%). This pyrosequencing allowed detection of higher than 85% of richness estimator Chao1 and Ace at the OTU_3% level. Higher coverage (≥0.97) and much less of rare types (singletons, only accounting for 24.5% of all OTU_3%) indicated that this pyrosequencing recovered most of bacteria inhabiting these biospheres. At the phylum level, the high relative sequence abundance (42.0% to 63.3%) showed that Proteobacteria was the dominant member at all these sampling sites. At the class level, Deltaproteobacteria, Gammaproteobacteria, and Flavobacteriia composed the majority of bacterial communities, and the relative abundance of Cyanobacteria and Bacilli varied significantly among the six samples. At the genus level, abundant OTUs related with sulfate reduction, including Desulfobulbus and Desulforhopalus, were identified. Shared and unique OTUs analysis revealed that, at the OTU_3% level, 508 OTUs were shared by all the six samples, and the number of unique OTUs ranged from 98 (R02) to 195 (NB04). Principal coordinates analysis PCoA analysis revealed that samples C04 and NB04 had the similar communities and were distinct from the others. Canonical correspondence analysis (CCA) revealed that temperature was the most significant factors that correlated with the bacterial community composition. The differences in bacterial compositions and diversities indicate that the similar sediment habitats contain a large variation in microbial biodiversity.     

双台子河口沉积物中细菌多样性分析

【目的】掌握双台子河口沉积物中细菌多样性及其群落结构的季节变化特征。【方法】于2009年4月、7月、10月和12月共4个航次在内陆河流入海口处进行四季样品的采集, 采用PCR-DGGE技术对沉积物中细菌多样性进行分析。【结果】通过序列比对发现, 该处沉积物中的细菌主要归属于5个细菌类群, 分别为变形菌门(52.6%)、放线菌门(15.8%)、拟杆菌门(10.5%)、酸杆菌门(5.3%)以及绿弯菌门(5.3%), 此外还有一部分分类地位尚不明确的细菌(10.5%)。在四季样品中变形菌门(52.6%)为优势菌群, 而在变形菌门中, δ亚群又占绝对优势地位。实验结果还显示四季沉积物中细菌Shannon-Wiener多样性指数范围为1.84?2.79, 且春夏两季沉积物中的Shannon-Wiener多样性指数比秋冬两季沉积物中Shannon-Wiener多样性指数大。【结论】双台子河口沉积物中的细菌多样性符合典型河口沉积物中细菌多样性的特征; 低温能导致沉积物中细菌多样性的减少。本研究为初步掌握双台子河口沉积物中细菌种类和组成状况提供了一定的参考, 同时也为该处海洋环境的监测及生物资源的保护提供了科学依据。