中国南海南沙海区沉积物中细菌16SrDNA多样性的初步研究

通过构建并分析中国南海南沙海区沉积物中细菌16S rRNA基因文库,对其遗传多样性进行了研究.发现沉积物中细菌16S rRNA基因主要来自变形细菌(Proteobacteria)的δ-,γ-,α-亚族和浮霉状菌目(Planctomycetales)等类群,所获的16S rRNA基因的序列与GenBank中已知的细菌16S rDNA序列差异较大,这一结果表明在中国南沙海区沉积物中存在丰富的微生物多样性,并潜藏着特有的微生物资源.     

中国对虾遗传多样性的RAPD分析

采用随机扩增多态DNA技术,对中国对虾的3个野生地理群体--朝鲜半岛西海岸产卵群体及其越冬群体、黄渤海中国沿岸群体和一个人工累代养殖群体进行了遗传多样性研究.用一组随机引物,对每个群体各20尾对虾的基因组DNA进行扩增,共获得110个重复性好且谱带清晰的RAPD标记,68.2%的标记在所有个体间保持稳定的一致性,表明中国对虾的遗传多样性水平偏低.4个群体的多态位点比例在20%～33.3%之间,群体内的遗传多态度为0.0093～0.0307.群体间遗传变异高于群体内.不同群体间遗传变异水平不同,朝鲜半岛西海岸产卵群体及其越冬群体相近且高于黄渤海沿岸群体,累代养殖群体遗传变异度最低.通过成对的遗传距离分析,用非加权的组平均法构建了上述4个群体中国对虾的谱系关系图,该聚类分析的结果与中国对虾在黄渤海的地理分布相吻合.     

南极阿德雷岛地表沉积物中细菌多样性及对环境的响应

提取南极阿德雷岛沉积物柱状样各层次的总DNA,利用PCR-RFLP方法,对沉积物中的细菌多样性及分布进行了研究,并探讨了它与环境的关系.结果表明,沉积物中的细菌多样性丰富,分属于8个类群,以CFB(Cytophaga-Flexibacter-Bacterioides)类群和Proteobacteria类群的β-,γ-亚群为主.在7 cm左右深度的沉积物中,可培养微生物和16 S rDNA序列多样性与其他深度的沉积物有明显不同,推测与环境的变化有关.同时还发现了大量与降解有机化合物相关的细菌,表明阿德雷岛的微生物生态系统已经受到了人类活动的影响.     

海洋沉积物中特有细菌类群的初步探讨

通过构建16S rRNA基因文库对中国南沙南海区沉积物中细菌多样性进行分析,并将获得16S rRNA基因序列与国际基因数据库GenBank进行相似性比较及聚类分析,结果发现来自海洋沉积物的未能培养的微生物往往组成一簇,并与现已获培养并鉴定的微生物自然分开,这一表象表明,海洋沉积物中存在特有的微生物属种。     

环境细菌遗传多样性分析的基因标签串测序法

把研究真核细胞表达基因种类和水平的基因标签串测序法引入环境细菌遗传多样性分析,选择细菌６Ｓ核糖体ＲＮＡ基因（ｒＤＮＡ）内０２６碱基对高变异区段为该基因变异类型的标签,连接成标答中并测序,用标签类型、频率和２多样性指数等参数在分子水平上分析环境细菌遗传多样性,建立了环境细菌遗传多样性分析的基因标签串测序法,该方法在分析环境隐含遗传多样性、提示污染生物学效应和评价环境质量等方面有的应用价值。因使用共同     

广西北部湾局部海域海洋沉积物细菌多样性及生物活性评估

为有效探究海洋新型天然微生物资源,本研究以广西北部湾局部海域海洋沉积物样品为研究对象,采用高通量测序技术和纯培养分离方法分析海洋沉积物样品中细菌群落组成及多样性,利用双层琼脂扩散法检测纯培养菌株的抗菌活性,通过PCR扩增法筛选基因组中含有7种功能基因的阳性菌株。高通量测序结果显示：海洋沉积物中检测到1407个操作分类单元（Operational Taxonomic Units,OTUs）;在门水平上,变形菌门（Proteobacteria）的物种丰度最高,其次是厚壁菌门（Firmicutes）和酸杆菌门（Acidobacteria）。纯培养分离纯化鉴定出278株菌,分布在3门5纲16目24科31属,其中YIM 150853为1个潜在新分离单元。生物活性评估实验表明,76.34%的纯培养菌株至少对1种指示菌有抑制作用,其中链霉菌YIM 150601、YIM 150634抑菌谱广、抑菌活性强,放线菌的功能基因检出率高于非放线菌。广西北部湾局部海域海洋沉积物中蕴藏着丰富的细菌资源,纯培养菌株具有抗菌及合成生物活性产物的潜力,是开发新型天然产物的新菌源。     

基于不同引物的湖泊沉积物中细菌群落结构多样性的比较研究

采用PCRD-DGGE技术,用两对引物对湖泊沉积物细菌群落结构进行了比较研究.结果表明:沉积物样品中细菌群落结构显示了明显的空间分布多样性差异,随着沉积深度的增加,细菌群落多样性指数降低.采用不同引物扩增的产物DGGE图谱聚类相近,均分成0～20cm和20～45cm两大类,但引物341f/534r导出的相似性Cs值在30.2%～75.9%之间,引物341f/926r得出的相似性Cs值在34.3%～84.1%之间,采用不同引物不同沉积深度样品群落结构之间的相似性和优势种动态变化存在差异.     

对虾养殖池底泥中光合细菌分离及其生物学特性

从虾池底泥样品富集分离出2株光合细菌,分别编号为M1和M2,均为革兰氏阴性菌,繁殖方式为二均分裂.细胞中含有细菌叶绿素a,经形态、生理生化特性初步鉴定,M1和M2菌株均为红假单胞菌属(Rhodopseudomonas)的成员,初步研究了NaCl浓度对其生长的影响.     

Genetic Diversity in Populations of Sepiella maindroni Using 16S rRNA Gene Sequence Analysis

Part of the 16S rRNA gene is amplified with PCR and sequenced for 5 populations of common Chinese cuttlefish Sepiella maindroni:three from the South China Sea,one from East China Sea and one from Japan.The result shows that a total of 5 nucleotide positions are found to have gaps or insertions of base pairs among these individuals,and 13 positions are examined to be variable in all the sequences,which range from 494 to 509 base pairs.All of the individuals are grouped into 7 haplotypes (h1-h7).No marked genetic difference is osberved among those populations.All of the individuals from Nagasaki belong to h1 and the h3 haplotype is found only in the coastal waters of China.A→←G transition in Nucleotide 255 is suggested to be taken as a kind of genetic marker to identify the populations distributed in East-South China Sea and the Nagasaki waters of Japan.     

墨吉对虾Penaeus merguiensis 线粒体16SrRNA 基因序列分析

比较墨吉对虾6个群体的线粒体16SrRNA基因约457个碱基对的DNA序列。结果表明这6个群体间的遗传距离（D）在0-0.0085之间，未发现广东的3个群体与澳洲或新加坡群体间存在地理群体以上的遗传差异。     

Community structure of β-Proteobacterial ammonia-oxidizing bacteria in prawn farm sediment

To examine the community structure of β-Proteobacterial ammonia-oxidizing bacteria （AOB） in prawn farm sediment, the 16S rRNA gene library was constructed with β-Proteobacterial AOB-specific primers. The library was screened by PCR-restriction fragment length polymorphism （RFLP） analysis and clones with unique RFLP patterns were sequenced. Two groups of β-Proteobacterial AOB, the Nitrosomonas and the Nitrosospira, were detected. The Nitrosomonas occupied an absolute dominant position, accounting for more than 90% of total clones in the clone library, while the Nitrosospira accounting for 5.48%. Nitrosomonas-affiliated clones were grouped into the Nitrosomonas marina and the Nitrosomonas sp. Nm143 clusters, and Nitrosospira-affiliated clones were grouped into the Nitrosospira cluster 1. No other clusters of β-Proteobacterial AOB were found. The results enriched our knowledge of AOB diversity in the prawn farm sediment and provided important foundational data for further functional studies of these microbes in mariculture environments.     

Community structure of β-Proteobacterial ammonia-oxidizing bacteria in prawn farm sediment

To examine the community structure of β-Proteobacterial ammonia-oxidizing bacteria (AOB) in prawn farm sediment, the 16S rRNA gene library was constructed with β-Proteobacterial AOB-specific primers. The library was screened by PCR-restriction fragment length polymorphism (RFLP) analysis and clones with unique RFLP patterns were sequenced. Two groups of β-Proteobacterial AOB, the Nitrosomonas and the Nitrosospira, were detected. The Nitrosomonas occupied an absolute dominant position, accounting for more than 90% of total clones in the clone library, while the Nitrosospira accounting for 5.48%. Nitrosomonas-affiliated clones were grouped into the N. marina and the N. sp. Nm143 clusters, and Nitrosospira-affiliated clones were grouped into the Nitrosospira cluster 1. No other clusters of β-Proteobacterial AOB were found. The results enriched our knowledge of AOB diversity in the prawn farm sediment and provided important foundational data for further functional studies of these microbes in mariculture environments.     

Community structure of β-Proteobacterial ammonia-oxidizing bacteria in prawn farm sediment

To examine the community structure of β-Proteobacterial ammonia-oxidizing bacteria (AOB) in prawn farm sediment, the 16S rRNA gene library was constructed with β-Proteobacterial AOB-specific primers. The library was screened by PCR-restriction fragment length polymorphism (RFLP) analysis and clones with unique RFLP patterns were sequenced. Two groups of β-Proteobacterial AOB, the Nitrosomonas and the Nitrosospira, were detected. The Nitrosomonas occupied an absolute dominant position, accounting for more than 90% of total clones in the clone library, while the Nitrosospira accounting for 5.48%. Nitrosomonas-affiliated clones were grouped into the Nitrosomonas marina and the Nitrosomonas sp. Nm 143 clusters, and Nitrosospira-affiliated clones were grouped into the Nitrosospira cluster 1. No other clusters of β-Proteobacterial AOB were found. The results enriched our knowledge of AOB diversity in the prawn farm sediment and provided important foundational data for further functional studies of these microbes in mariculture environments.     

云台山白云岩表层土可培养细菌多样性

运用纯培养法和基于16S rRNA基因序列的系统发育分析贵州云台山白云岩表层土可培养细菌的多样性.稀释涂布平板分离共获得131株细菌,限制性内切酶HhaⅠ和HaeⅢ对扩增的16S rRNA基因片段进行酶切分型,根据ARDRA酶切图谱划分为40个操作分类单元.16S rRNA基因序列测定和系统发育分析结果显示,这些菌株隶属于包括厚壁菌门(Firmicutes,64.89%)、β-变形菌纲(β-Proterbacteria,3.82%)、γ-变形菌纲(γ-Proterbacteria,17.56%)、放线细菌门(Actinobacteria,11.45%)和拟杆菌门(Bacteroidetes,3.82%)等5大类群,共13个属.优势菌群为芽胞杆菌属(Bacillus,53.44%),亚优势菌群为寡养单胞菌属(Stenotrophomonas,9.16%)、假单胞菌属(Pseudomonas,8.40%)和微小杆菌属(Exiguobacterium,8.40%).15.00%的有效序列与GeneBank已知序列相似性小于等于97%,可能为潜在新类群.研究表明,云台山白云岩喀斯特地区不仅含有较为丰富的细菌物种多样性,且潜藏着许多新的微生物资源.     

鄱阳湖流域底泥微生物对环境变量的响应

以鄱阳湖流域4个支流共6个地点(信江XS1、信江下水湾XS3、饶河RS、饶河内湖RHS、修水SS、赣江GS)的底泥样品为研究对象,利用16 S rRNA基因扩增技术探究底泥微生物对环境变量的响应.结果显示:6个采样点的底泥样品的微生物群落结构大致分为4类; 底泥的pH值、TOC、TN、TP和重金属含量对微生物群落结构及微生物多样性存在着显著的影响.4条河流底泥微生物多样性指数顺序为赣江>信江>修水>饶河,同时发现饶河的Cu、Zn含量在4条河流中最高,这说明高Cu、Zn会对微生物多样性产生负面的影响; 河流与其所对应湖区底泥样品中微生物多样性指数比较发现:饶河>饶河内湖,信江下水湾>信江,这种结果顺序的不一致主要是由底泥中营养物质含量不同和某些重金属元素胁迫导致的.     

海洋细菌的分子鉴定分类

从南海海底沉积物中分离到一批细菌,克隆了其中产抗菌活性物且培养形态多变的细菌ＺＳ１１０的１６ＳｒＤＮＡ,通过和比较１６ＳｒＤＮＡ的部分序列,对ＺＳ１１０进行了分子鉴定,结果表明它是一株适应了海洋环境的芽孢杆菌属的枯草芽孢杆菌种。     

西沙野生诺尼内生细菌群落多样性初步研究

采用构建16S rDNA克隆文库方法,首次对我国海南永兴岛西沙野生诺尼果内生细菌的群落结构和多样性进行初步研究.首次采用Mothur软件对诺尼果内生细菌克隆文库数据进行分析,构建Mothur软件数据分析平台.构建的西沙野生诺尼果内生细菌16S rDNA克隆文库中,93个克隆分属于12个不同可操作分类单元(operational taxonomic units,OTU),序列比对结果表明大部分克隆与已知细菌16S rDNA序列相似性较高.分别属于变形菌门(Proteobacteria)的Alpha、Beta、Gamma亚群,放线菌门(Actinobacteria)和厚壁菌门(Firmicutes).水库杆菌属(Piscinibacter sp.)细菌为野生诺尼果内生细菌优势菌群.优势菌属分别为水库杆菌属(Piscinibacter sp.)为80.65%,蛋白胨菌属(Peptoniphilus sp.)和链球菌属(Streptococcus sp.)均为3.23%,甲基杆菌属(Methylobacterium sp.)为2.15%.研究结果表明西沙野生诺尼果中存在较为丰富的内生细菌资源,将为进一步研究奠定理论基础.