西南印度洋深海热液硫化物区沉积物微生物群落结构和固氮基因多样性

A sediment sample was collected from a deep-sea hydrothermal vent field located at a depth of 2 951 m on the Southwest Indian Ridge. Phylogenetic analyses were performed on the prokaryotic community using polymerase chain reaction（PCR） amplification of the 16 S rRNA and nifH genes. Within the Archaea, the dominant clones were from marine benthic group E（MBGE） and marine group I（MGI） belonging to the phyla Euryarchaeota and Thaumarchaeota, respectively. More than half of the bacterial clones belonged to the Proteobacteria, and most fell within the Gammaproteobacteria. No epsilonproteobacterial sequence was observed. Additional phyla were detected including the Actinobacteria, Bacteroidetes, Planctomycetes, Acidobacteria, Nitrospirae, Chloroflexi, Chlorobi, Chlamydiae, Verrucomicrobia, and candidate divisions OD1, OP11, WS3 and TM6, confirming their existence in hydrothermal vent environments. The detection of nifH gene suggests that biological nitrogen fixation may occur in the hydrothermal vent field of the Southwest Indian Ridge. Phylogenetic analysis indicated that only Clusters I and III NifH were present. This is consistent with the phylogenetic analysis of the microbial 16 S rRNA genes, indicating that Bacteria play the main role in nitrogen fixation in this hydrothermal vent environment.     

东太平洋海隆深海热液区沉积物古菌多样性分析

采用PCR-RFLP方法对东太平洋海隆深海热液区3个站位沉积物中的古菌多样性进行了初步研究.结果显示,从古菌16S rRNA基因文库中随机挑取的296个阳性克隆分属奇古菌门（Thaumarchaeota,47.64%）、广古菌门（Euryarchaeota,44.93%）、泉古菌门（Crenarchaeota,6.77%）和未分类古菌（0.68%）,其中优势菌群为奇古菌门的亚硝化侏儒菌属（Nitrosopumilus,35.47%）和广古菌门的热原体纲（Thermoplasmata,27.03%）,DHVE3、DHVE5、DHVE6、MBGB和MBGE类群在沉积物样品中也均有发现.另外,3个站位沉积物中古菌类群组成存在差异,S5-TVG1站位样品文库的97个古菌克隆分属奇古菌门（49.48%）、广古菌门（49.48%）和泉古菌门（1.03%）,S14-TVG10站位样品文库的103个古菌克隆由奇古菌门（84.47%）和广古菌门（15.53%）组成,S16-TVG12站位样品文库的96个古菌克隆包括广古菌门（71.88%）、泉古菌门（19.79%）、奇古菌门（6.25%）和未分类古菌（2.08%）.研究结果表明,东太平洋海隆深海热液区沉积物中古菌多样性丰富,存在着许多新的古菌菌群;不同站位古菌菌群结构以及多样性存在差异,这与其所处环境的热液活动密切相关.     

Colonization of Sulfurovum sp. on the gill surfaces of Alvinocaris longirostris, a deep-sea hydrothermal vent shrimp

Deep‐sea hydrothermal vents are unique light‐independent ecosystems that are sustained by chemosynthetic bacteria. For many of the invertebrates inhabiting in such environments, bacteria play essential roles in both energy acquisition and detoxification of potentially toxic gases such as H₂S. In this study, the bacterial flora present on the gills of Alvinocaris longirostris (Bresiliidae: Caridea), a shrimp inhabiting hydrothermal vents (1532 m depth) at the Hatoma Knoll of the Okinawa Trough, was investigated. Bacterial 16S rDNA fragments were successfully amplified from the gills and 70% of these fragments showed an identical pattern in the restriction fragment length polymorphism analysis. These fragments were assigned to the ribotype AL‐1. Phylogenetic analyses suggest that AL‐1 forms a monophyletic clade with Sulfurovum spp. (ε‐Proteobacteria). Fluorescence in situ hybridization for AL‐1 and electron microscopy showed the presence of short‐rod bacteria lining up on the cuticular layer of the surface of the gill filaments. These results suggest that bacterial association with gills also occurs in bresiliid shrimps.     

Spatial distribution, diversity and composition of bacterial communities in sub-seafloor fluids at a deep-sea hydrothermal field of the Suiyo Seamount

Spatial distribution, diversity, and composition of bacterial communities within the shallow sub-seafloor at the deep-sea hydrothermal field of the Suiyo Seamount, Izu-Bonin Arc, Western Pacific Ocean, were investigated. Fluids were sampled from four boreholes in this area. Each borehole was located near or away from active vents, the distance ranging 2–40 m from active vents. In addition, fluids discharging from a natural vent and ambient seawater were sampled in this area. We extracted DNA from each sample, amplified bacterial 16S rRNA genes by PCR, cloned the PCR products and sequenced. The total number of clones analyzed was 348. Most of the detected phylotypes were affiliated with the phylum Proteobacteria, of which the detection frequency in each clone library ranged from 84.6% to 100%. The bacterial community diversity and composition were different between hydrothermal fluids and seawater, between fluids from the boreholes and the vent, and even among fluids from each borehole. The relative abundances of the phylotypes related to Thiomicrospira, Methylobacterium and Sphingomonas were significantly different among fluids from each borehole. The phylotypes related to Thiomicrospira and Alcanivorax were detected in all of the boreholes and vent samples. Our findings provide insights into bacterial communities in the shallow sub-seafloor environments at active deep-sea hydrothermal vent fields.     

南海西沙海槽沉积物柱MD05-2902细菌多样性

A sediment core MD05-2902 was collected from the deep-sea basin of the Xisha Trough. The vertical distribution and diversity of bacteria in the core was investigated through ten sub-sampling with an interval of 1 m using bacterial 16 S rRNA gene as a phylogenetic bio-marker. Eighteen phylogenetic groups were identified from 16 S rRNA gene clone libraries. The dominant bacterial groups were JS1, Planctomycetes and Chloroflexi, which accounted for 30.6%, 16.6%, and 15.6% of bacterial clones in the libraries, respectively. In order to reveal the relationship between biotic and abiotic data, a nonmetric multidimensional scaling analysis was performed. The result revealed that the δ15N, δ13C, total organic carbon and total organic nitrogen possibly influenced the bacterial community structure. This study expanded our knowledge of the biogeochemical cycling in the Xisha Trough sediment.     

Phylogenetic diversity of planktonic bacteria in the Chukchi Borderland region in summer

Planktonic bacteria are abundant in the Chukchi Borderland region. However, little is known about their diversity and the roles of various bacteria in the ocean. Seawater samples were collected from two stations K2S and K4S where sea ice was melting obviously. The analysis of water samples with fluorescence in situ hybridization (FISH) showed that DMSP-degrading bacteria accounted for 13% of the total bacteria at the station K2S. No aerobic anoxygenic phototrophic (AAP) bacteria were detected in both samples. The bacterial communities were characterized by two 16S rRNA gene clone libraries. Sequences fell into four major lineages of the domain Bacteria, including Proteobacteria (Alpha, Beta and Gamma subclasses), Bacteroidetes, Actinobacteria and Firmicutes. No significant difference was found between the two clone libraries. SAR11 and Rhodobacteraceae clades of Alphaproteobacteria and Pseudoalteromonas of Gammapro-teobacteria constituted three dominant fractions in the clone libraries. A total of 191 heterotrophic bacterial strains were isolated and 76% showed extracellular proteolytic activity. Phylogenetic analysis reveals that the isolates fell into Gammaproteobacteria, Bacteroidetes, Actinobacteria and Firmicutes. The most common genus in both the bacterial isolates and protease-producing bacteria was Pseudoalteromonas. UniFrac data showed suggestive differences in bacterial communities between the Chukchi Borderland and the northern Bering Sea.     

劳盆地热液喷口沉积物微生物多样性初步研究

采用聚合酶链式反应限制性片段长度多态性分析(PCR-RFLP)技术对劳盆地热液区TVG9站位沉积物中细菌、古菌多样性进行了调查,并构建其细菌、古菌种群的16S rRNA基因文库.研究结果表明,该站位细菌包含变形杆菌(Proteobacteria)、硝化螺旋菌(Nitrospira),疣微菌(Verrucomicrobia)等3个类群,其中变形杆菌占据优势地位,由Alphaproteobactria,Gamaproteobactria,Deltaproteobactria等3个亚群组成.古菌包含泉古菌(Crenarchaeota)和广古菌(Euryarchaeota),其中泉古菌占优势,由MG Ⅰ (Marine Group Ⅰ)和MCG(Miscellaneous Crenarchaeotal Group)两类群组成,而广古菌主要由MBGE(Marine Benthic Group E)组成.旨在揭示深海热液区的微生物多样性,为生态环境的研究提供理论支持.     

渤海沉积物中细菌群落结构和基因丰度的空间分布特征

This study investigated differences in the community structure and environmental responses of the bacterial community in sediments of the Bohai Sea.Illumina high-throughput sequencing technology and real-time PCR were used to assay the bacterial 16S rRNA genes in the surface sediments of 13 sampling stations in the Bohai Sea.The results showed that sediments at the majority of the 13 sampling stations were contaminated by heavy metal mercury.The main phyla of bacteria recorded included Proteobacteria(52.92%),Bacteroidetes(11.76%),Planctomycetes(7.39%),Acidobacteria(6.53%)and Chloroflexi(4.97%).The genus with the highest relative abundance was Desulfobulbus(4.99%),which was the dominant genus at most sampling stations,followed by Lutimonas and Halioglobus.The main factors influencing bacterial community structure were total organic carbon,followed by depth and total phosphorus.The content of lead,cadmium,chromium,copper and zinc had a consistent effect on community structure.Arsenic showed a negative correlation with bacterial community structure in most samples,while the impact of mercury on community structure was not significant.The bacterial community in sediment samples from the Bohai Sea was rich in diversity and displayed an increase in diversity from high to low latitudes.The data indicated that the Bohai Sea had abundant microbial resources and was rich in bacteria with the potential to metabolize many types of pollutants.     

黄海西北近岸沉积物中细菌群落空间分布特征

采用16S rDNA文库和变性梯度凝胶电泳(DGGE)技术,对黄海西北部3个近岸站位沉积物中细菌群落多样性及空间分布特征进行了调查和解析。对表层沉积物16S rDNA序列统计表明,各站位细菌群落多样性很高,γ-和δ-变形菌纲分别占克隆序列总数的20%~32%,是沉积物中的绝对优势类群。DGGE图谱分析表明,同一站位中不同深度的细菌群落结构相似性较高,而不同站位间群落结构相差较远。研究表明在黄海西北近岸沉积物中细菌群落多样性较高,优势类群明显,在较小尺度范围内群落结构的垂直变化不明显。     

西菲律宾海比科尔陆架深海沉积物古菌多样性研究

通过构建16S rRNA 基因文库, 对西太平洋西菲律宾海东板比科尔陆架5 个不同层位沉积物样品中的古菌的多样性进行了研究, 并获得了465 个有效克隆63 个OTUs （Operational Taxonomic Units）。通过16S rRNA 序列与GenBank 已知序列的同源性比较及构建系统进化树的结果显示...     

Peripheral communities of the Eastern Lau Spreading Center and Valu Fa Ridge: community composition,temporal change and comparison to near‐vent communities

Western Pacific hydrothermal vents will soon be subjected to deep‐sea mining and peripheral sites are considered the most practical targets. The limited information on community dynamics and temporal change in these communities makes it difficult to anticipate the impact of mining activities and recovery trajectories. We studied community composition of peripheral communities along a cline in hydrothermal chemistry on the Eastern Lau Spreading Center and Valu Fa Ridge (ELSC‐VFR) and also studied patterns of temporal change. Peripheral communities located in the northern vent fields of the ELSC‐VFR are significantly different from those in the southern vent fields. Higher abundances of zoanthids and anemones were found in northern peripheral sites and the symbiont‐containing mussel Bathymodiolus brevior, brisingid seastars and polynoids were only present in the northern peripheral sites. By contrast, certain faunal groups were seen only in the southern peripheral sites, such as lollipop sponges, pycnogonids and ophiuroids. Taxonomic richness of the peripheral communities was similar to that of active vent communities, due to the presence of non‐vent endemic species that balanced the absence of species found in areas of active venting. The communities present at waning active sites resemble those of peripheral sites, indicating that peripheral species can colonize previously active vent sites in addition to settling in the periphery of areas of venting. Growth and mortality were observed in a number of the normally slow‐growing cladorhizid stick sponges, indicating that these animals may exhibit life history strategies in the vicinity of vents that differ from those previously recorded. A novel facultative association between polynoids and anemones is proposed based on their correlated distributions.     

Depth matters: Microbial eukaryote diversity and community structure in the eastern North Pacific revealed through environmental gene libraries

Protistan community structure was examined from 6 depths (1.5, 20, 42, 150, 500, 880 m) at a coastal ocean site in the San Pedro Channel, California. A total of 856 partial length 18S rDNA protistan sequences from the six clone libraries were analyzed to characterize diversity present at each depth. The sequences were grouped into a total of 259 Operational Taxonomic Units (OTUs) that were inferred using an automated OTU calling program that formed OTUs with approximately species-level distinction (95% sequence similarity). Most OTUs (194 out of 259) were observed at only one specific depth, and only two were present in clone libraries from all depths. OTUs were obtained from 21 major protistan taxonomic groups determined by their closest BLAST matches to identified protists in the NCBI database. Approximately 74% of the detected OTUs belonged to the Chromalveolates, with Group II alveolates making up the largest single group. Protistan assemblages at euphotic depths (1.5, 20 and 42 m) were characterized by the presence of clades that contained phototrophic species (stramenopiles, chlorophytes and haptophytes) as well as consumers (especially ciliates). Assemblages in the lower water column (150, 500 and 800 m) were distinct from communities at shallow depths because of strong contributions from taxa belonging to euglenozoans, acantharians, polycystines and Taxopodida (Sticholonche spp. and close relatives). Species richness (Chao I estimate) and diversity (Shannon index) were highest within the euphotic zone and at 150 m, and lowest for protistan assemblages located in the oxygen minimum zone (500 and 880 m). Multivariate analyses (Bray-Curtis coefficient) confirmed that protistan assemblage composition differed significantly when samples were grouped into shallow (≤150 m) and deep water assemblages (≥150 m).     

Microbial diversity in the sediments of the southern Mariana Trench

Microbial diversity in the abyssal sediments beneath the seafloor of 30,94,and 151cm near the southern end of the Mariana Trench was analyzed in the Illumina HiSeq 2500 platform.Results show that the microbial populations were dominated by bacteria but merely no archaea were identifi ed at the three depths.In the bacterial community,Proteobacteria and Firmicutes dominated the total taxon tags,followed by Bacteroidetes,Actinobacteria,Planctomycetes,Cyanobacteria,and Chloroflexi,which together account for over 99%of the total population.Similar to that in the seawater in the trench,the operational taxonomic units(OTUs)belonging to Gammaproteobacteria from the sediment samples showed high abundance.However,common bacterial OTUs in the water of the trench including Nitrospirae and Marinimicrobia were hardly found in the sediments from the southern Mariana Trench or the hadal region.Therefore,this study documented for the first time the compositions of microbial diversity in the trench sediments,revealed the difference in microbial diversity in water and sediment of the trench and will enrich the knowledge on the microbial diversity in the abyssal areas.     

印度洋深海热液区可培养细菌的分子鉴定与系统发育分析

采用MMJHS寡营养培养基从印度洋深海热液区沉积物和热液硫化物中分离获得16株细菌,通过16S rDNA序列比对和生理生化分析,对它们进行了鉴定并构建了系统发育树.结果表明,12株细菌属于γ-变型菌(γ-Gammaproteobacteria),其中6株属于盐单胞菌属(Halomonas),4株属于嗜冷杆菌属(Psychrobacter),2株属于食碱菌属(Alcanivorax);其余4株属于芽孢杆菌(Bacillus),其中1株为地衣芽孢杆菌(Bacillus licheniformis).分离获得的16株细菌中,4株为革兰氏阳性细菌,12株为革兰氏阴性细菌;H2S反应、吲哚测定反应、M.R和V-P均呈阴性.本研究为深入认识和开发利用深海热液区微生物资源奠定了基础.     

冲绳海槽南部热液区附近柱状沉积物中微生物的垂直分布多样性及其对环境因子的响应

近年来,海底热液环境中的微生物及其环境适应机制已经成为海洋科学研究的热点。目前,相关的研究主要集中在表层沉积物及微生物的水平分布多样性方面,而对柱状沉积物中微生物垂直分布多样性研究却很少。本文基于西太平洋冲绳海槽南部热液区附近S2站位的柱状沉积物样品,通过对其不同层位的样品进行分离培养和16S rRNA基因高通量测序,揭示了样品中可培养微生物和总体微生物的垂直群落分布特征,同时结合对样品主量元素、微量元素、碳氮含量等指标的评估和冗余分析等统计学方法,讨论了微生物群落结构及其对环境因子的响应。研究发现该位点的柱状沉积物有机质含量较为贫乏,存在两个富含Cu-Zn-Pb的层;各个层位的沉积物中微生物类群均以变形菌为主要类群,同时表层沉积物表现出更高的微生物多样性。此外研究还表明柱状沉积物中有机碳含量与其微生物的群落组成有着更为密切的关系。总之, 本研究的结果和获得的菌种资源为进一步深入研究海底热液环境中微生物参与元素地球化学循环的过程提供了一定的基础。     

北极深海沉积物中细菌和古菌群落结构研究

在北极深海沉积物生态系统中,微生物的群落结构由有机质输入、能量的可用性及其他环境因素决定.然而,全球气候变暖及其导致的冰盖提前融化正在影响微生物的多样性.为描述北极深海沉积物中的微生物群落结构及其与环境因素的相关性,我们利用罗氏454对北极深海沉积物样品的16S rDNA扩增子进行了测序,对细菌和古菌群落的丰富度、成分、结构及其系统发育分类地位进行了描述.硫还原和化能有机营养类是细菌群落中的主要类群;而古细菌群落主要是由微生物的关系最为密切的氨氧化奇古菌门(96.66%)和产甲烷古生菌界(3.21%).这项研究描述了北极极点附近深海沉积物(> 3500米)中的微生物多样性,将为以后研究类似环境中微生物代谢过程和途径等功能分析奠定基础.     

海底热液喷口流体中H2S浓度数据统计及其探测技术进展

统计了全球38个热液活动区264个热液喷口流体样品中的H_2S浓度数据,结果表明H_2S浓度主要受岩浆去气、水岩反应、相分离作用影响,而广泛使用的非气密保压采样技术不能反映原始喷口流体状态和化学组成,可能会造成H_2S浓度测量误差。为了提高测量数据的精度,一方面需要进一步发展气密保压采样技术,以提高样品的保真水平;另一方面利用深海原位电化学传感器或拉曼光谱系统进行海底原位探测也将是一个重要的发展方向。