祁连山天然气水合物赋存区钻孔细菌多样性

分析了青海省祁连山冻土区天然气水合物赋存区DK-6钻孔的4个样品,对岩心样品进行处理提取微生物总基因组,采用PCR构建了细菌16S rDNA基因文库,4个文库包括44个OTU,其中有厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)、变形杆菌门(Proteobacteria)(包括α-、β-和y-变形杆菌亚群)、放线菌纲(Actinobacteria)和异常球菌-栖热菌门(Deinococcus-Thermus)5类,煤、泥岩、粉砂岩等不同岩性的微生物群落之间显示出较大的组成差异,优势菌不同.细菌菌群多样性随采集点地质环境不同而有较明显的变化,天然气水合物含量、水含量、有机质含量等环境因素对冻土区天然气水合物赋存区中细菌菌群有一定的影响.4个样品中存在的微生物大部分可以代谢有机烃类,在天然气水合物环境的特殊条件下,外界环境因素制约了微生物的种类.     

不同培养条件对海洋沉积环境细菌的选择性分离

海洋环境中存在着大量未被培养和利用的微生物资源。本研究对一份南海沉积物样品采用不同培养温度、盐度、pH、样品稀释倍数和营养浓度条件进行可培养细菌的多样性研究。经过16S rRNA基因序列分析,获得825株菌分属于5个门,8个纲,17个目,26个科,57个属。分离到最多的属级类群为芽胞杆菌属（Bacillus）。通过不同培养条件的分离实验发现,厚壁菌类群在4 °C~60 °C、0%~15%盐度、pH 5~8及不同的样品稀释倍数和营养浓度实验条件中均为优势培养类群,具有广泛的环境适应性,但2~200的样品稀释倍数可以大大减少分离培养基中厚壁菌的数量。放线菌类群在4 °C低温和0%NaCl添加条件下的可培养多样性较高,同时pH 6和寡营养培养基有助于分离获得稀有放线菌类群。另外,本研究发现新物种资源的获取几率分别在寡营养培养基、5%~10%较高盐度和60°C高温培养有所增加。分离获得的5个主要细菌门类分别为厚壁菌门（Firmicutes,86%）、放线菌门（Actinobacteria,13%）、变形菌门（Proteobacteria,1%）、拟杆菌门（Bacteroidetes）和异常球菌-栖热菌门（Deinococcus-Thermus）。本研究共分离得到29株潜在新种,分别属于芽胞杆菌纲（Bacilli）、放线菌纲（Actinobacteria）、酸微菌纲（Acidimicrobiia）、嗜热油菌纲（Thermoleophilia）、红色杆菌纲（Rubrobacteria）和异常球菌纲（Deinococci）。海洋环境微生物新类群、海洋放线菌稀有类群等微生物新资源的选择性分离培养提供了有效的方法和方案,为后期的深入开展打下良好的基础。     

造纸废水活性污泥的微生物群落结构及多样性分析

为探究造纸废水活性污泥中微生物群落结构多样性以及对造纸废水处理效果的影响，利用Illumina MiSeq 高通量测序方法，分析在处理造纸废水过程中，同一运行阶段两个并联氧化沟内活性污泥的微生物群落与多样性组成。结果表明，系统中处理造纸废水的活性污泥在同一废水条件下微生物群落结构总体稳定，优势细菌为绿弯菌门(Chloroflexi)、拟杆菌门（Bacteroidota）、变形菌门（Proteobacteria）、Myxococcota、放线菌门(Actinobacteria）、厚壁菌门(Firmicutes）等。最重要的优势细菌类群为Chloroflexi，相对丰度占比为47.67%~48.22%，远远高于其他废水中Chloroflexi的占比，其中厌氧绳菌纲（Anaerolineae）是其主要成员，占比84.39%~88.34%，可针对性地去除造纸废水中的污染物。造纸废水活性污泥样品中存在大量特殊功能菌群，其在废水中污染物尤其是木质素的去除中发挥着重要作用。     

多囊卵巢综合征患者与健康人群肠道菌群差异性分析

目的比较多囊卵巢综合征(PCOS)患者与健康人群间肠道菌群的差异,为后续研究提供参考。方法采用16S rDNA扩增子测序法对30例PCOS患者(PCOS组)和20例健康人(健康组)粪便标本中菌群结构进行分析,并比较两组之间的区别。结果与健康组相比,PCOS组患者肠道菌群α多样性降低。PCOS组患者肠道厚壁菌门(Firmicutes)、放线菌门(Actinobacteria)丰度高于健康组,拟杆菌门(Bacteroidetes)丰度低于健康组(均P0.05)。两组对象肠道梭菌门(Clostridia)、放线菌门(Actinobacteria)丰度差异无统计学意义(均P0.05)。PCOS组患者肠道变形杆菌纲(Proteobacteria)、芽胞杆菌纲(Bacilli)、红椿杆菌纲(Coriobacteriia)丰度高于健康组,拟杆菌纲(Bacteroidia)丰度低于健康组(均P0.05)。PCOS组患者肠道布劳特菌属(Blautia)、霍氏真杆菌属(Eubacterium_hallii_group)、阴沟杆菌属(Agathobacter)丰度高于健康组,拟杆菌属(Bacteroides)、粪杆菌属(Faecalibacterium)丰度低于健康组(均P0.05)。两组对象肠道双歧杆菌属(Bifidobacterium)、埃希菌-志贺菌属(Escherichia-Shigella)丰度差异无统计学意义(均P0.05)。结论 PCOS患者存在肠道菌群失调情况,但其是否是PCOS发展的潜在致病因素需要进一步研究。     

性激素变化对小鼠肠道菌群构成的影响

肠道菌群在维持宿主免疫和消化系统功能方面发挥着重要作用,肠道菌群的多样性和丰富度是衡量宿主健康状况的重要生理指标。性激素对动物生长发育发挥重要作用,但其对肠道菌群构成影响的相关实验研究相对较少。本研究以模式生物小鼠（Mus musculus）为对象,探究性激素的变化对小鼠肠道菌群构成的影响。采用外科手术方式建立小鼠去势模型,通过16S r RNA高通量测序技术,研究性激素对小鼠肠道菌群构成的影响。研究结果表明,雌性小鼠和雄性小鼠去势后,性激素水平显著下降。肠道菌群在门水平上,正常小鼠和去势小鼠肠道细菌群落均由拟杆菌门（Bacteroidetes）、厚壁菌门（Firmicutes）、变形菌门（Proteobacteria）、埃普西隆杆菌门（Epsilonbacteraeota）、髌骨细菌门（Patescibacteria）、放线菌门（Actinobacteria）、软壁菌门（Tenericutes）、脱铁杆菌门（Deferribacteres）、酸杆菌门（Acidobacteria）和蓝藻细菌门（Cyanobacteria）组成,主要菌群为厚壁菌门和拟杆菌门物种,两门占物种相对丰度百分比...     

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。     

光唇鱼仔稚幼鱼肠道菌群与养殖水体细菌群落的相关性

采用16S rRNA高通量测序,系统研究了光唇鱼(Acrossocheilus fasciatus)仔鱼、稚鱼和幼鱼肠道菌群组成及与同时期养殖水体细菌群落的相关性。研究结果表明,光唇鱼仔鱼、稚鱼和幼鱼的肠道菌群中Chao1指数和Shannon指数均没有显著变化(P>0.05);而随着光唇鱼幼体的发育,养殖水体中Chao1指数和Shannon指数呈现显著下降趋势(P<0.05)。光唇鱼仔鱼和稚鱼肠道菌群中的优势菌门由变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)组成,而同时期养殖水体中优势菌门为变形菌门;光唇鱼幼鱼肠道菌群中的优势菌门为梭杆菌门(Fusobacteria)和变形菌门,同时期养殖水体中优势菌门由变形菌门、拟杆菌门和梭杆菌门组成。线性回归分析结果显示,随光唇鱼幼体发育,在光唇鱼肠道菌群中变形菌门、厚壁菌门和梭杆菌门相对丰度的时序变化趋势与其在养殖水体中相同。在属水平上,光唇鱼仔鱼、稚鱼肠道菌群中优势菌属均为醋酸杆菌属(Acetobacter),而幼鱼肠道菌群中优势菌属为鲸杆菌属(Cetobacte...     

柴达木盆地尕斯库勒盐湖区邻近水体对湖泊微生物群落的贡献

【目的】探究尕斯库勒盐湖生态系统中邻近水体对湖泊微生物的贡献。【方法】采集尕斯库勒盐湖区湖水、沉积物以及邻近的泉水、河水和盐田的水样,对其进行地球化学分析;通过16S rRNA基因的Illumina MiSeq高通量测序分析样品的微生物群落组成。【结果】尕斯库勒盐湖区水体和沉积物中的优势门是变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)和广古菌门(Euryarchaeota)。盐度和pH是影响尕斯库勒盐湖区群落组成的最主要环境因素。邻近水体对湖泊水体和沉积物的贡献分别为12.94%和7.53%。【结论】邻近水体对尕斯库勒盐湖微生物群落的贡献有限。     

圈养条件下新疆马鹿两个亚种的肠道菌群比较分析

肠道菌群组成复杂，与宿主肠道内环境动态平衡和健康息息相关。马鹿是鹿科动物中分布最广的种类，亚种分化众多，但现有马鹿肠道菌群研究较少。为丰富马鹿肠道菌群数据，以马鹿的两个亚种为阶元，运用高通量16S rRNA基因测序技术检测了7头雄性天山马鹿和7头雄性塔里木马鹿的肠道微生物，对其核心菌群、结构组成、多样性及肠道微生物的差异进行分析，旨在了解不同马鹿亚种肠道微生物组成差异，为两亚种间肠道菌群的差异提供参考。研究结果表明，塔里木马鹿菌群多样性及丰度低于天山马鹿。厚壁菌门（Firmicutes）、放线菌门（Actinobacteria）、变形菌门（Proteobacteria）及拟杆菌门（Bacteroidetes）为两组马鹿的优势菌门。此外，天山马鹿肠道中分解蛋白质、脂质的拟杆菌门和变形菌门相对丰度高于塔里木马鹿，而分解植物纤维的双歧杆菌（Bifidobacterium）、瘤胃球菌科UCG 014（Ruminococcaceae UCG 014）的相对丰度低于塔里木马鹿。本研究对天山马鹿及塔里木马鹿肠道菌群组成进行初步了解，可根据研究结果调整其饲料配比，为马鹿饲养管理的改善提供参考。     

福州左海湖冬春季浮游细菌群落16S rRNA多样性

【目的】为揭示福州左海湖冬春不同季节水体中浮游细菌群落的多样性。【方法】通过对2012年1月(冬季)和2012年4月(春季)的左海湖水体样品构建细菌16S rRNA基因克隆文库,对2个季节样品中细菌群落的多样性和均匀度等进行比较,并对其群落结构特征进行分析。【结果】冬季左海细菌群落的多样性指数Shannon-Wiener(H)达到3.53,均匀度指数(E)达到0.66;春季细菌群落的多样性指数Shannon-Wiener(H)为3.37,均匀度指数(E)为0.64,这2个季节样品的细菌群落均具有很高的多样性,但均匀度都偏低。2个季节水体中共检测到5个细菌门类:α-变形菌纲(α-proteobacteria)、β-变形菌纲(β-proteobacteria)、拟杆菌门(bacteroidetes)、放线菌门(actinobacteria)、蓝细菌门(cyanobacteria),此外还存在一些未被认知的序列。冬季的优势菌群为β-proteobacteria,次优势菌群为cyanobacteria;春季的最优势菌群为cyanobacteria。【结论】左海湖水体细菌在冬春2季节中的菌群结构存在差异,特别是最优势的细菌发生了很大变化,其中冬季细菌群落的多样性及均匀度相对较高。     

Bacterial diversity in the sediment from polymetallic nodule fields of the Clarion-Clipperton Fracture Zone

The Clarion-Clipperton Fracture Zone (CCFZ) is located in the northeastern equatorial Pacific and contains abundant polymetallic nodules. To investigate its bacterial diversity, four libraries of 16S rRNA genes were constructed from sediments of four stations in different areas of the CCFZ. In total, 313 clones sequenced from the 4 libraries were assigned into 14 phylogenetic groups and 1 group of 28 unclassified bacteria. High bacterial diversity was predicted by the rarefaction analysis. The most dominant group overall was Proteobacteria, but there was variation in each library: Gammaproteobacteria was the most dominant group in two libraries, E2005-01 and ES0502, while Alphaproteobacteria and Deltaproteobacteria were the most dominant groups in libraries EP2005-03 and WS0505, respectively. Seven groups, including Alphaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Betaproteobacteria, Acidobacteria, Actinobacteria, and Bacteroidetes, were common to all four libraries. The remaining minor groups were distributed in libraries with different patterns. Most clones sequenced in this study were clustered with uncultured bacteria obtained from the environment, such as the ocean crust and marine sediment, but only distantly related to isolates. Bacteria involved in the cycling of metals, sulfur and nitrogen were detected, and their relationship with their habitat was discussed. This study sheds light on the bacterial communities associated with polymetallic nodules in the CCFZ and provides primary data on the bacterial diversity of this area.     

Bacterial community composition and diversity of a full-scale integrated fixed-film activated sludge system as investigated by pyrosequencing

The integrated fixed-film activated sludge (IFAS) system is a variation of the activated sludge wastewater treatment process, in which hybrid suspended and attached biomass is used to treat wastewater. Although the function and performance of the IFAS system are well studied, little is known about its microbial community structure. In this study, the composition and diversity of the bacterial community of suspended and attached biomass samples were investigated in a full-scale IFAS system using a highthroughput pyrosequencing technology. Distinct bacterial community compositions were examined for each sample and appeared to be important for its features different from conventional activated sludge processes. The abundant bacterial groups were Betaproteobacteria (59.3%), Gammaproteobacteria (8.1%), Bacteroidetes (5.2%), Alphaproteobacteria (3.9%), and Actinobacteria (3.2%) in the suspended sample, whereas Actinobacteria (14.6%), Firmicutes (13.6%), Bacteroidetes (11.6%), Betaproteobacteria (9.9%), Gammaproteobacteria (9.25%), and Alphaproteobacteria (7.4%) were major bacterial groups in the attached sample. Regarding the diversity, totals of 3,034 and 1,451 operational taxonomic units were identified at the 3% cutoff for the suspended and attached samples, respectively. Rank abundance and community analyses demonstrated that most of the diversity was originated from rare species in the samples. Taken together, the information obtained in this study will be a base for further studies relating to the microbial community structure and function of the IFAS system.     

Effect of the Earthworms Lumbricus terrestris and Aporrectodea caliginosa on Bacterial Diversity in Soil

Earthworms ingest large amounts of soil and have the potential to radically alter the biomass, activity, and structure of the soil microbial community. In this study, the diversity of eight bacterial groups from fresh soil, gut, and casts of the earthworms Lumbricus terrestris and Aporrectodea caliginosa were studied by single-strand conformation polymorphism (SSCP) analysis using both newly designed 16S rRNA gene-specific primer sets targeting Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Bacteroidetes, Verrucomicrobia, Planctomycetes, and Firmicutes and a conventional universal primer set for SSCP, with RNA and DNA as templates. In parallel, the study of the relative abundance of these taxonomic groups in the same samples was performed using fluorescence in situ hybridization. Bacteroidetes, Alphaproteobacteria, and Betaproteobacteria were predominant in communities from the soil and worm cast samples. Representatives of classes Flavobacteria and Sphingobacteria (Bacteroidetes) and Pseudomonas spp. (low-abundant Gammaproteobacteria) were detected in soil and worm cast samples with conventional and taxon-targeting SSCP and through the sequence analysis of 16S rRNA clone libraries. Physiologically active unclassified Sphingomonadaceae (Alphaproteobacteria) and Alcaligenes spp. (Betaproteobacteria) also maintained their diversities during transit through the earthworm intestine and were found on taxon-targeting SSCP profiles from the soil and worm cast samples. In conclusion, our results suggest that some specific bacterial taxonomic groups maintain their diversity and even increase their relative numbers during transit through the gastrointestinal tract of earthworms.     

Diversity and structure of bacterial communities in Fildes Peninsula, King George Island

To examine the bacterial community structure in the Fildes Peninsula, King George Island, Antarctica, we examined the bacterial diversity and community composition of samples collected from lacustrine sediment, marine sediment, penguin ornithogenic sediments, and soils using culture-dependent and culture-independent methods. The 70 strains fell into five groups: Actinobacteria, Bacteroidetes, Firmicutes, Gammaproteobacteria, and Betaproteobacteria. Bacterial diversity at the phylum level detected in Denaturing Gradient Gel Electrophoresis (DGGE) profiles comprised Proteobacteria (including the subphyla Alpha-, Beta-, Gamma-, Deltaproteobacteria), Bacteroidetes, Firmicutes, Chlorobi, and Deinococcus-Thermus. Gammaproteobacteria was identified to be the dominant bacterial subphylum by cultivation and DGGE method. By cluster analysis, the overall structure and composition of bacterial communities in the soil and lacustrine sediment were similar to one another but significantly different from bacterial communities in penguin ornithogenic sediment and marine sediment, which were similar to one another. The majority of 16S rDNA sequences from cultured bacteria were closely related to sequences found in cold environments. In contrast, a minority of 16S rDNA sequences from the DGGE approach were closely related to sequences found in cold environments.     

Microbial composition of biofilms in a brewery investigated by fatty acid analysis,fluorescence in situ hybridisation and isolation techniques

Biofilms associated with brewery plants can harbour spoiling microorganisms that potentially damage the final product. Most beer-spoiling microorganisms are thought to depend on numerous interactions with the accompanying microbiota. However, there is no information on the microbial community structure of biofilms from bottling plants. The conveyors that transport the bottles to and from the plant are known as potential sources of microbial contamination of beer. Consequently, the material buildup from two conveyors was analysed using a cultivation/isolation approach, and the culture-independent techniques of whole cell fatty acid analysis and fluorescence in situ hybridisation (FISH). Heterogeneous communities were present at both conveyors. Although characteristic fatty acids for Eukarya were present, FISH-signals for Eukarya were extremely low. The Proteobacteria, in particular the Gammaproteobacteria, were abundant at both sample sites. Bacterial isolates were obtained for every dominating group detected by FISH: the Alphaproteobacteria, Betaproteobacteria and Gammaproteobacteria, the Xanthomonadaceae, the Actinobacteria, the Bacteroidetes and the Firmicutes.     

基于PCR-DGGE指纹的南海海绵共附生细菌优势种群的揭示与系统发育分析

采用PCR-DGGE指纹、克隆测序和系统发育分析技术较系统地对我国南海贪婪倔海绵(Dysidea avara)和澳大利亚厚皮海绵(Craniella australiensis)共附生的优势细菌进行了研究。研究发现变形菌门(Proteobacteria)细菌是这两种海绵中的主要优势细菌,贪婪倔海绵中的变形菌包含了α、β、γ三种类型,而澳大利亚厚皮海绵中仅有γ一种类型。两种海绵都有拟杆菌(Bacteroidetes),但是具体的种类不同。这些细菌都是第一次在海绵中被发现。澳大利亚厚皮海绵共附生的优势细菌还包括放线菌属(Actinobacterium)及厚壁菌门(Firmicutes)细菌,菌群多样性要比贪婪倔海绵的丰富。两种海绵尽管来自于同一海域但其共附生优势细菌的组成明显不同,这说明海绵共附生微生物具有宿主特异性。     

Phylogenetic diversity, composition and distribution of bacterioplankton community in the Dongjiang River, China

Bacterioplankton community compositions in the Dongjiang River were characterized using denaturing gradient gel electrophoresis (DGGE) and 16S rRNA gene clone library construction. Water samples in nine different sites were taken along the mainstem and three tributaries. In total, 24 bands from DGGE gels and 406 clones from the libraries were selected and sequenced, subsequently analyzed for the bacterial diversity and composition of those microbial communities. Bacterial 16S rRNA gene sequences from freshwater bacteria exhibited board phylogenetic diversity, including sequences representing the Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Actinobacteria, Bacteriodetes, Verrucomicrobia, and candidate division TM7. Members of Betaproteobacteria group were the most dominant in all sampling sites, followed by Gammaproteobacteria, Alphaproteobacteria, and Actinobacteria. DGGE profiles and the ∫-LIBSHUFF analysis revealed similar patterns of bacterial diversity among most sampling sites, while spatial distribution variances existed in all sites along the river basin. Statistical analysis showed that bacterial species distribution strongly correlated with environmental variables, such as nitrate and ammonia, suggesting that nitrogen nutrients may shape the microbial community structure and composition in the Dongjiang River. This study had important implications for the comparison with other rivers elsewhere and contributed to the growing data set on the factors that structure bacterial communities in freshwater ecosystems.     

Bacterial, archaeal and eukaryotic diversity of smooth and pustular microbial mat communities in the hypersaline lagoon of Shark Bay

The bacterial, archaeal and eukaryotic populations of nonlithifying mats with pustular and smooth morphology from Hamelin Pool, Shark Bay were characterised using small subunit rRNA gene analysis and microbial isolation. A highly diverse bacterial population was detected for each mat, with 16S rDNA clones related to Actinobacteria, Bacteroidetes, Chloroflexi, Cyanobacteria, Gemmatimonas, Planctomycetes, Alphaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Verrucomicrobia and candidate division TM6 present in each mat. Spirochaetes were detected in the smooth mat only, whereas candidate division OP11 was only detected in the pustular mat. Targeting populations with specific primers revealed additional cyanobacterial diversity. The archaeal population of the pustular mat was comprised purely of Halobacteriales, whereas the smooth mat contained 16S rDNA clones from the Halobacteriales, two groups of Euryarchaea with no close characterised matches, and the Thaumarchaea. Nematodes and fungi were present in each mat type, with diatom 18S rDNA clones only obtained from the smooth mat, and tardigrade and microalgae clones only retrieved from the pustular mat. Cultured isolates belonged to the Firmicutes, Gammaproteobacteria, Alphaproteobacteria, Bacteroidetes, Actinobacteria, Cyanobacteria, and Halobacteriales. The mat populations were significantly more diverse than those previously reported for Hamelin Pool stromatolites, suggesting specific microbial populations may be associated with the nonlithifying and lithifying microbial communities of Hamelin Pool.     

Phylogenetic diversity of microorganisms associated with the deep-water sponge Baikalospongia intermedia

The diversity of bacteria associated with deep-water sponge Baikalospongia intermedia was evaluated by sequence analysis of 16S rRNA genes from two sponge samples collected in Lake Baikal from depths of 550 and 1204 m. A total of 64 operational taxonomic units, belonging to nine bacterial phyla, Proteobacteria (classes Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, and Deltaproteobacteria), Actinobacteria, Planctomycetes, Cloroflexi, Verrucomicrobia, Acidobacteria, Chlorobi, and Nitrospirae, including candidate phylum WS5, were identified. Phylogenetic analysis showed that the examined communities contained phylotypes exhibiting homology to uncultured bacteria from different lake ecosystems, freshwater sediments, soil and geological formations. Moreover, a number of phylotypes were relative to psychrophilic, methane-oxidizing, sulfate-reducing bacteria, and to microorganisms resistant to the influence of heavy metals. It is noted that the unusual habitation conditions of deep-water sponges contribute to the taxonomic diversity of associated bacteria and have an influence on the presence of functionally important microorganisms in bacterial communities.     

九龙江河口区nirS型反硝化细菌多样性及系统发育学分析

【目的】结合16S rRNA基因克隆文库和nirS基因克隆文库的分析,揭示九龙江河口区nirS型反硝化细菌多样性。【方法】选取九龙江河口区一富营养化采样点,分别采集水样及沉积物样品,进行理化因子的测定并提取细菌总DNA。以水样DNA构建16S rRNA基因克隆文库,以沉积物DNA构建nirS基因克隆文库,分析微生物群落结构的多样性并构建系统发育树。【结果】从16S rRNA基因克隆文库中获得86条有效序列,按97%的序列相似性划分为53个OTU,分别属于Proteobacteria门、Planctomycetes门、Bacteroidetes门、Actinobacteria门、Firmicutes门和Chloroflexi门。其中属于Proteobacteria门OTU的克隆子占克隆数的62.9%,是最优势的类群,分属于Alphaproteobacteria、Betaproteobacteria、Gammaproteobacteria和Deltaproteobacteria纲等。从nirS基因克隆文库中获得190条有效序列,翻译为氨基酸序列后,按82%的序列相似性划分为60个OTU,并定位到属的水平。其中Proteobacteria门是最优势的类群,占文库克隆子总数的71.6%,包括Alphaproteobacteria纲(5.8%)、Betaproteobacteria纲(49.0%)和Gammaproteobacteria纲(16.9%)。nirS基因克隆文库中丰度最高的OTU与GenBank中的一株可培养反硝化菌Thauera sp. R-26906具有100%的序列相似性。【结论】九龙江河口区的微生物以及亚硝酸盐还原酶基因(nirS)具有丰富的多样性。大部分NirS序列在GenBank中的最相似序列来源于河口、海湾等相似的环境。