西北印度洋卡尔斯伯格脊卧蚕热液羽流影响区细菌群落结构特征及其演化

【目的】热液羽流影响区包括热液羽流流经区域和羽流中性浮力层下方受热液颗粒物影响的区域。随着热液羽流的演化,热液羽流影响区内微生物群落的结构组成也会发生相应的变化,但是,由于观测和取样困难等原因,迄今热液羽流影响区不同空间位置微生物的群落结构特征及其在月际尺度上的演化尚不清楚。【方法】中国大洋49航次在卧蚕1号热液喷口东南侧300 m处投放了沉积物捕获器锚系,在不同离底高度开展了为期18个月的观测和时序采水。本文采用Illumina MiSeq高通量测序技术对水样中的微生物类群进行测序分析,结合现场实时探测的浊度异常资料,研究卧蚕热液区附近中性浮力羽流和热液颗粒沉降区细菌群落结构的特征和演化及其影响因素。【结果】结果表明,样品中细菌群落以γ-变形菌纲(Gammaproteobacteria)、弯曲菌纲(Camplylobacteria)、α-变形菌纲(Alphaproteobacteria)、拟杆菌纲(Bacteroidia)、梭菌纲(Clostridia)和脱硫叶菌纲(Desulfobulbia)为主。在时间上,优势类群的相对丰度随浊度起伏发生变化,当浊度异常值升高时,弯曲菌纲相对丰度...     

西南印度洋中脊热液羽状流中微生物化石的发现及意义

深海热液微生物因其生存于海底极端环境,能够提供深部生物圈和生物地球化学信息而成为当今海洋科学领域关心的热点问题之一.本文以西南印度洋中脊热液羽状流水体中悬浮颗粒物为研究对象,选取典型的3个CTD采水站位,对离底42-500m的不同水层悬浮体过滤的滤膜进行扫描电镜观察和X射线能谱面扫描分析.结果表明,热液羽状流颗粒物中含有大量的微生物化石,包括丝状、球状、杆状和簇状等4种形态.对不同形态微生物化石能谱分析表明Si、O、Ca、Fe和Zn等元素在微生物化石中都有高值出现,其中丝状体中具有Ti和Mn的高值出现,球状体中Ba具有高值,说明微生物化石化过程中不同类型的微生物体对金属元素的选择性吸收,钙化和铁硅酸盐岩化是微生物化石化的主要机制.微生物化石在羽状流中的分布状况能够提供活动热液喷口的空间信息,为我们寻找喷口提供帮助.     

西北印度洋卡尔斯伯格脊卧蚕热液羽流影响区古菌群落动态变化特征

【目的】热液羽流在浮力上升和非浮力侧向迁移过程中与背景海水不断进行物质和能量的交换。因海底热液活动和洋流的流速流向均存在动态变化性,热液羽流的高度、物理化学参数和微生物群落也随时空发生演化。然而,由于缺乏热液羽流的长期监测和时序取样,微生物群落(尤其是古菌)的多样性及其时空演化尚不清楚。【方法】2018年7月-2019年6月,在卧蚕1号热液喷口东南300 m处放置了一套带有2个沉积物捕获器(分别距海底300 m和40 m)和一个浊度仪(距海底150 m)的锚系潜标,对热液羽流和热液羽流下方颗粒物沉降区的近底水样进行了为期18个月的观测和时序采样。本研究采用16S rRNA基因高通量测序技术研究了水样中古菌群落的多样性,以了解羽流层和近底沉降层古菌群落结构的特征与时空演化。【结果】热原体纲(Thermoplasmata)、甲烷八叠球菌纲(Methanosarcinia)和甲烷杆菌纲(Methanobacteria)等热液来源古菌,当浊度异常值较高时,它们的丰度也增加。从空间上看,热液羽流区和近底沉降区古菌群落结构在门和纲水平上具有相似性,但在目水平上表现出差异性,而且各主要类群的相对丰度也存在差异,其中热原体纲在羽流层中的丰度普遍高于近底沉降层,而氨氧化古菌和甲烷八叠球菌纲在沉降层的丰度较高。【结论】卧蚕热液区东南300 m热液羽流层和近底沉降层均受到了热液的影响,羽流层受热液影响的程度相对更显著,而近底沉降层除了受到一定程度的热液影响外,还可能受到了再悬浮沉积物的影响。热液贡献大小的动态变化和底层沉积物再悬浮可能是造成热液区近端水体古菌群落时空异质性的主要因素。本研究深化了对热液影响区古生菌群落结构及其月际尺度时空分布特征的认识。     

宁波港压载水的细菌多样性研究

为揭示宁波港压载水的微生物群落结构在人类活动影响下的分布差异及对环境因子变化的响应趋势。该课题从压载水中分离细菌,通过微生物自动鉴定系统(Sherlock MIS)、分子生物学16S rRNA基因的PCR扩增及RFLP技术,对宁波港压载水的细菌结构特征进行比对和分析,并通过16S rRNA基因文库解析研究压载水中微生物群落多样性。结果表明印度洋水样中存在假交替单胞菌属(Pseudoalteromonas sp.)、交替单胞菌属(Alteromonas sp.)、海杆菌属(Marinobacter sp.)和玫瑰杆菌属(Roseobacter sp.)、肠杆菌属(Enterobacter sp.);新加坡水样中存在人苍白杆菌属(Ochrobactrum sp.)、甲基杆菌属(Methylobacterium sp.)、鞘鞍醇单胞菌(Sphingomonas sp.)、嗜冷杆菌属(Psychrobacter sp.)、假单胞菌(Pseudomonas sp.)、伯克霍尔德氏菌(Burkholderia sp.)和葡萄球菌属(Staphylococcus sp.)假单胞菌属(Pseudomonas sp.);而美洲水样中存在弧菌属(Vibrio sp.)、军团菌属(Legionella sp.)、莫拉氏菌属(Moraxella sp.)和不动杆菌属(Acinetobacter sp.)。压载水中含有大量的潜在致病菌。     

宜宾浓香型白酒酿造过程中可培养细菌的系统发育多样性

【目的】为较系统地了解宜宾浓香型白酒酿造过程中可培养细菌的多样性,得到一些潜在的微生物资源。【方法】采用改良的NA培养基和高氏I号培养基分离、去除冗余,测定所得细菌纯培养物的16S rRNA基因,进行系统发育分析。【结果】分离得到603株细菌,4株菌的序列与GenBank中典型菌株序列相似性低于97%,代表着潜在新类群;599株菌与GenBank中34个属、101个种的典型菌株序列相似性大于97%,其中以Bacillus为绝对优势菌(315株),Streptomyces(121株)、Lysinibacillus(35株)、Staphylococcus(45株)为次优势菌,其余各属菌株均在10株以下。而且有16个属均只检测到1株菌。【结论】宜宾浓香型白酒发酵过程中的细菌呈现出较为丰富的多样性和一定的稳定性。     

利用改良培养基探究西太平洋海水可培养细菌多样性

[目的]西太平洋复杂的海洋生态环境孕育了其独特的生物群落,蕴含着种类丰富的海洋微生物资源.本研究基于分离培养技术探究了西太平洋海域不同水深细菌的多样性,并尝试通过改良培养基提高海洋细菌可培养性.[方法]采用改良的2216E固体培养基(IMA)、R2A固体培养基(R2A)、MBM固体培养基(MBM)、TCBS固体培养基(...     

山西左云县采煤区人工湿地冬季沉积物细菌群落多样性

【背景】人工湿地处理煤炭矿井废水效果显著,而关于人工湿地沉积物的细菌群落结构和特征研究较少。【目的】阐明人工湿地净化采煤区废水的效应和细菌群落结构及特征。【方法】对山西省左云县采煤区人工湿地不同采样点水质进行监测,利用高通量测序技术对人工湿地沉积物中细菌16S rRNA基因V3-V4区进行测序,分析沉积物中细菌群落组成。【结果】采煤区废水流经人工湿地后水质得到有效改善,其中生化需氧量(Biochemical oxygen demand,BOD5)、化学需氧量(Chemical oxygen demand,CODCr)、总氮(Total nitrogen,TN)、总磷(Total phosphorus,TP)去除率分别达到76.2%、93.4%、73.4%和99.3%;测序共获得2 832个操作分类单元(Operational taxonomic units,OTUs),共753个属,分布于51个门的150个纲;4个采样点中优势菌门为变形菌门(Proteobacteria,50%-64.7%)和拟杆菌门(Bacteroidetes,15.8%-21.2%),变形菌门中优势菌纲为β-变形菌纲和γ-变形菌纲,其中M1、M2、M3中富含氨氮氧化细菌的β-变形菌纲和γ-变形菌纲丰度最高;M4沉积物细菌OTU最多,多样性最高;沉积物特有优势菌属Sulfurimonas、Sulfuricurvum的相对丰度与NH4+-N的含量显著负相关,Azoarcus菌属相对丰度与TN存在显著正相关,Novosphingobium菌属相对丰度与TP含量存在显著正相关。【结论】煤矿废水流经人工湿地可得到有效的净化,细菌多样性在人工湿地生态功能中起到了决定作用。     

九香虫(椿象)体内可培养细菌的多样性分析

【目的】认识药用昆虫九香虫(Aspongopus chinesis Dallas)成虫体内可培养细菌资源多样性。【方法】运用纯培养法、反转录重复因子扩增(BOXA1R-PCR)分析技术、16S r RNA基因测序和系统发育分析对样品中可培养细菌进行多样性研究,测定了分离菌株的抗菌特性、吲哚乙酸(IAA)含量和产淀粉酶活性等指标。【结果】通过6种不同培养基共分离得到52株菌落特征不同的细菌菌株。基于菌落特征和BOXA1R-PCR图谱选取12株代表菌株用于16S r RNA基因序列测定。16S r RNA基因序列系统发育分析显示,52株菌株分属于芽孢杆菌属(Bacillus)、假单胞菌属(Pseudomonas)、寡养单胞菌属(Stenotrophomonas)和伯克霍尔德氏菌属(Burkholderia)4个属,其中芽孢杆菌属(Bacillus)为优势菌属。分离到的52株细菌有44株(占总分离菌株的84.6%)表现出对供试病原菌具有较好的抑制作用,高达94.2%的分离菌株能产IAA,有43株(占总分离菌株的82.7%)表现出淀粉酶活性。【结论】九香虫内细菌种群较为多样,具有潜在应用价值。     

河南鲁山五大温泉水细菌多样性分析

【背景】位于河南中部的平顶山市含有丰富的温泉资源,对温泉水中的微生物多样性和群落结构进行研究有助于更好地开发利用温泉资源。【目的】探究河南鲁山五大温泉水中(上汤、中汤、下汤、温汤和神汤)细菌多样性。【方法】采用Illumina Hi Seq2500 PE250技术对鲁山五大温泉水细菌的16S r RNA基因V3-V4变异区序列进行测序,应用UPARSE、Mothur、Silva、MUSCLE和QIIME等软件整理和统计样品序列数目和操作分类单元(Operational taxonomic unit,OTU)数量,最后分析五大温泉水中细菌的丰度和多样性。【结果】上汤、中汤、下汤、温汤和神汤分别获得56 017、68 319、65 247、59 340、68 825条有效Tags,聚类为670、287、337、598、381个OTU。细菌分类分析表明,五大温泉水的细菌超过40个门,变形菌门(Proteobacteria)占84.12%,是优势菌;在属分类阶元上,五大温泉水的细菌超过239个属,上汤的优势菌是不动杆菌属(Acinetobacter),中汤的优势菌是Tepidimonas,下汤的优势菌是Vogesella和不动杆菌属(Acinetobacter),温汤的优势菌是Sphingopyxis和新鞘脂菌属(Novosphingobium),神汤的优势菌是Aquabacterium。【结论】鲁山五大温泉水中含有丰富的微生物资源,为后续五大温泉水中的微生物资源开发奠定了基础。     

云南兰坪铅锌矿区细菌多样性研究

目的研究云南省兰坪铅锌矿区矿石样品中的细菌群落结构。方法构建细菌16S rRNA高变区基因文库。结果矿石内部细菌种类丰富多样,文库测序结果表明矿石样品中细菌分属于4个门,27个属;其中变形菌门（Proteobacteria）类群是优势种群,占所有细菌总克隆数的51.7%,而且大多数属于γ-变形菌纲亚群。各克隆序列分析结果表明,其中部分细菌序列与已报道物种的相似性较低。结论表明云南省兰坪铅锌矿矿石样品中可能存在新的细菌种类。     

Bacterial diversity in deep-sea sediment from northeastern Pacific Ocean

16S rDNA sequencing method is one of the effectively used culture-independent techniques in recent years. In this study, 16S rDNA sequencing method was used to investigate the bacterial diversity in deep-sea sediment from northeastern Pacific polymetallic nodule province. Total DNAs were extracted by using 2 different methods (chemical method and DNA extracting kit method). After purification, genomic DNA was amplified by using 2 universal primers (27F and 1492R). Clones were selected and sequenced randomly. After the sequences were checked by using the Chimera Check Program of the RDP database, a bacterial 16S rRNA gene library of 79 clones was established. Phylogenetic analysis indicated that 79 clones could be divided into 11 phylotypes. Gamma Proteobacteria (22.8%) and alpha Proteobacteria (16.5%) were the dominant components of the sediment bacterial community, followed by 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%) and uncultured bacteria (17.7%). Gamma Proteobacteria also dominated at slices 0–2 cm and 4–6 cm. Different slices had different types of bacteria, alpha Proteobacteria, gamma Proteobacteria, delta Proteobacteria, Planctomycetacia, Nitrospira, Actinobacteria and Acidobacteria, however, appeared in all slices. Pseudomonas is common in many different deep-sea environments. In this study, it accounted for 22.2% of the total gamma Proteobacteria.     

Bacterial group II introns in a deep-sea hydrothermal vent environment

Group II introns are catalytic RNAs and mobile retrotransposable elements known to be present in the genomes of some nonmarine bacteria and eukaryotic organelles. Here we report the discovery of group II introns in a bacterial mat sample collected from a deep-sea hydrothermal vent near 9 degrees N on the East Pacific Rise. One of the introns was shown to self-splice in vitro. This is the first example of marine bacterial introns from molecular population structure studies of microorganisms that live in the proximity of hydrothermal vents. These types of mobile genetic elements may prove useful in improving our understanding of bacterial genome evolution and may serve as valuable markers in comparative studies of bacterial communities.     

印度洋深海沉积物石油烃降解菌分离、鉴定与多样性分析

烃类物质在海洋环境中广泛分布,深海可能含有特殊的烃降解微生物。本研究通过对西南印度洋中脊与印度洋中部深海沉积物中石油降解菌的富集培养和分离鉴定,从6个站点的样品中共分离得到800株菌,通过BOX-PCR去重复菌株后,对其中183株菌进行了16S rRNA基因序列分析,发现这些菌分属于23个属;其中,γ-变形菌纲的食烷菌属(Alcanivorax)和放线菌纲的微杆菌属(Microbacterium)占优势。此外,还发现了食烷菌属2个潜在的新种、假海栖菌属(Pseudooceanicola)1个潜在新种。高通量测序结果证明,富集菌群中γ-变形菌纲是优势菌,主要包括食烷菌属、盐单胞菌属(Halomonas)、海杆菌属(Marinobacter)等。结合可培养菌与高通量测序结果,食烷菌属、盐单胞菌属、海杆菌属、交替假单胞菌(Pseudoalteromonas)、海源菌属(Idiomarina)与微杆菌属(Microbacterium)是沉积物样品中常见的石油烃降解菌,迪茨氏菌属(Dietzia)、红球菌属(Rhodococcus),假单胞菌属(Pseudomonas)、赤杆菌属(Erythrobacter)与副球菌属(Paracoccus)等可能也参与了烃的降解。     

Microbial diversity of a sulphide spire located in the Edmond deep-sea hydrothermal vent field on the Central Indian Ridge

A culture‐independent molecular phylogenetic survey was carried out for a bacterial and archaeal community of a mineralized crust coating a sulphide spire, which was collected from the Edmond vent field (23° S, 69° E, 3300 m depth) on the Central Indian Ridge. Small‐subunit rRNA genes (16S rDNA) were amplified from environmental DNA by PCR utilizing Bacteria‐specific, and Archaea‐specific 16S rDNA primers. PCR products were cloned and 26 bacterial and nine archaeal unique sequence types (phylotypes) were identified from 150 clones analysed by restriction fragment length polymorphism, representing eight and four distinct lineages, respectively. The majority (>90%) of the bacterial phylotypes group with the ?‐Proteobacteria and confirms the global prevalence of ?‐Proteobacteria in deep‐sea hydrothermal environments. Among the ?‐Proteobacteria, >40% of the phylotypes were closely related to the recently isolated deep‐sea vent thermophilic chemolithoautotrophic sulphur‐reducer, Nautilia lithotrophica. A single bacterial sequence was nearly identical (99% similarity) to the thermophilic hydrogen‐oxidizing Hydrogenobacter thermolithotrophum, and is the first report of Hydrogenobacter at deep‐sea hydrothermal vents. A majority (97%) of the archaeal phylotypes grouped with the ‘Deep‐sea Hydrothermal Vent Euryarchaeotal Group’, a phylogenetic lineage of uncultured Archaea that have only been reported from other deep‐sea hydrothermal vents on the Mid‐Atlantic Ridge, East Pacific Rise, Juan de Fuca Ridge, Isu–Ogasawara Arc, Okinawa Trough and the Manus Basin. A single sequence was closely related to the hyperthermophilic sulphur‐reducing Thermococcales frequently found in diverse deep‐sea vent environments. Scanning electron micrographs of the mineralized crust reveal abundant filamentous, rod and coccoidal forms encased in sulphur and sulphide mineral precipitate, suggesting that the thermophilic chemolithoautorophs and sulphide‐producing heterotrophs may influence the architecture and sulphur cycling of the sulphide spire.     

Archaeal diversity and community development in deep-sea hydrothermal vents

Over the past 35 years, researchers have explored deep-sea hydrothermal vent environments around the globe and studied a number of archaea, their unique metabolic and physiological properties, and their vast phylogenetic diversity. Although the pace of discovery of new archaeal taxa, phylotypes and phenotypes in deep-sea hydrothermal vents has slowed recently, bioinformatics and interdisciplinary geochemistry-microbiology approaches are providing new information on the diversity and community composition of archaea living in deep-sea vents. Recent investigations have revealed that archaea could have originated and dispersed from ancestral communities endemic to hydrothermal vents into other biomes on Earth, and the community structure and productivity of chemolithotrophic archaea are controlled primarily by variations in the geochemical composition of hydrothermal fluids.     

Bacterial diversity in deep-sea sediments from different depths

Seven sediment samples have been examined, taken from different depths of the deep-sea in the range of 1159m to 6482m. A total of 75 different 16S rDNA sequences (149 clones) analyzed clustered into the Proteobacteria, Gram-positive bacteria, Cytophaga, Planctomyces, and Actinomycetes and many sequences were from microorganisms that showed no phylogenetic affiliation with known bacteria. Clones identical to 16S rDNA sequences of members of the genus Pseudomonas were observed in all of the sediments examined. The second group of common sequences cloned from six sediment samples was related to the 16S rDNA sequence of a chemoautotrophic bacterium, the Solemya velum symbiont. Five 16S rDNA sequences from three sediments were related to those of the Alvinella pompejana epibiont which is a member of the -Proteobacteria. Only one sequence was obtained that was closely related to the 16S rDNA of the barophilic bacterium, Shewanella benthica, which might be a minor population in the deeper sediments. -Proteobacteria-related sequences were cloned from sediments obtained from sites near man-made garbage deposits and a Calyptogena community. These environments obviously would be richer in nutrients than other sites, and might be expected to show more types of bacteria than other deep-sea sediments. A large number of cloned sequences in this study showed very low identity to known sequences. These sequences may represent communities of as-yet-uncultivated microorganisms in the sediments.