深海热液喷口微生物群落研究进展

1977年在太平洋上的加拉帕戈斯群岛附近深2 500 m的深海热液区首次发现了独立的生命体系,它被认为是海洋生物学研究领域中最为重大的发现之一[1-2]。自此,对深海热液喷口生物群落的调查研究吸引了无数科学家的眼球,成为了海洋生物学研究领域的热点之一。30余年来,随着海洋生物学研     

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.     

东海陆架表层沉积物微生物多样性初步研究

提要从东海陆架DH-13站点的表层沉积物中提取环境基因组DNA,通过PCR和TA克隆构建了细菌和古菌的16SrDNA基因文库,并对克隆子文库进行系统发育分析。结果表明:细菌序列以变形菌门(Proteobacteria,41.5%)居多,其次是浮霉菌门(Planctomycetes,10.9%)、放线菌门(Actino-bacteria,8.9%)和CFBgroup(7.9%),另外还有少量酸杆菌门(Acidobacteria)、疣微菌门(Verru-comicrobia)、硝化螺旋菌门(Nitrospira)和厚壁菌门(Firmicutes)等;古菌序列全部来自泉古生菌门(Crenarchaeota),其中Marine Crenarchaeotic Group Ⅰ(MGⅠ,93.6%)是绝对优势菌,还有少量序列属于Miscellaneous Crenarchaeotic Group(MCG)、Marine BenthicG roup C(MBGC)和Marine Benthic Group A(MBGA)。文库多样性分析结果显示东海陆架表层沉积物中有着丰富多样的微生物群落,细菌的多样性更为显著。     

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.     

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。     

厚壳贻贝(Mytilus coruscus)养殖区沉积物微生物多样性及固碳潜力研究

海洋蓝碳是海洋碳汇研究的重要领域,厘清不同蓝碳生境中沉积物有机碳组分格局是当前研究的热点之一。为更好地理解此问题,现以近海厚壳贻贝养殖区这一特殊蓝碳生境为对象,解析沉积物中的碳氮组分格局;进一步通过关联沉积物微生物群落、结合卡尔文循环和还原三羧酸循环的关键基因相对丰度分析,评估厚壳贻贝养殖区沉积物的固碳潜力。结果表明,相较于非养殖区,厚壳贻贝养殖区沉积物惰性碳的累积较大,氮组分主要以氨氮形式存在;同时养殖区高微生物量碳和微生物量氮指示了其沉积物中碳周转较快,碳氮组分特征差异明显。沉积物微生物高通量测序结果显示养殖区沉积物微生物主要以Gamma变形菌纲和Delta变形菌纲为主,且微生物类群与颗粒有机碳、惰性碳等碳组分存在明显的相关性。与惰性碳存在明显正相关关系的硫微螺菌科(Thiomicrospiraceae)丰度在养殖区沉积物中显著高于非养殖区沉积物。贻贝养殖区沉积物包含cbbL在内的6种关键功能基因,固碳潜力明显。研究结果将为进一步探究蓝碳生境的有机碳来源和微生物固碳效率提供基础依据。     

印尼3个地区热泉中可培养细菌的分子鉴定与系统发育分析

采用MSM寡营养培养基从印度尼西亚Pantai cermin,Kalianda和Banyu wedang地区的热泉水样、泥样以及沉积物中根据菌落的形态学特征共分离获得69株细菌,通过菌株的16S rDNA的限制性片段长度多态性(PCR-RFLP)分析、序列比对和生理生化分析,对其进行鉴定并构建系统发育树.结果表明,分离...     

军曹鱼的分子遗传特性研究

采用随机扩增多态性DNA(RAPD)和线粒体DNA(mtDNA)的限制性片段长度多态性(RFLP)分析方法对广东湛江近海军曹鱼Rachycentron canadum的分子遗传学特征进行了研究。RAPD研究结果显示17个引物共检测到119个位点,其中多态位点比例P为80．85％,遗传相似系数S为0．7400,遗传距离D为0．2600,Nei基因多样性指数H为0．3009,Shannon信息指数I为0．4498。结果表明,军曹鱼的遗传多样性比较丰富。用19种识别5、6碱基的限制性内切酶对军曹鱼的mtDNA RFLP进行了分析研究,构建其mtDNA物理图谱;用引物5’-GTG ATC TGA AAA ACC ACC GTT G-3’和5’-AAT AGG AAG TAT CAT TGC GGT TTG ATG-3’扩增线粒体细胞色素b区段,得到稳定的850bp大小的特异性条带,用18种识别4-6碱基的限制性内切酶对扩增片段的限制性长度多态性进行分析,并测定其序列。     

南极普里兹湾及邻近海域沉积物微生物多样性与生理生化研究

对中国第29次南极科学考察采自南极普里兹湾及邻近海域25个站位的海洋沉积物样品进行了细菌、真菌的分离培养,经细菌16SrRNA、真菌ITS基因序列分析及系统发育分析,共鉴定得到12个属的28株细菌和7个属的10株真菌。细菌API 20NE生理生化及真菌产胞外酶活性实验显示,分离培养的细菌和真菌多数可产生水解酶类,初步认定其在参与微生物体内新陈代谢、适应南极极端环境方面发挥作用。这些南极低温条件下产生的酶类有可能在工业生产中有一定的应用价值。本研究丰富了对南极普里兹湾及邻近海域沉积物微生物多样性的认识,筛选获得了一批具有产酶特性的低温微生物可为进一步的应用研究奠定基础。     

帘蛤科4种养殖蛤群体遗传多样性和种间关系的fAFLP分析

利用荧光标记扩增片段长度多态性(fAFLP)技术对文蛤(Meretrix meretrix)、青蛤(Cyclina sinensis)、菲律宾蛤仔(Ruditapes philippinarum)和硬壳蛤(Mercenaria mercenaria)4种帘蛤科贝类的群体遗传多样性和种间关系进行了研究。选择EcoRⅠ/MseⅠ进行酶切,使用6个E 3/M 3引物组合进行扩增,共获得1 096个位点,多态位点比率95.1%,片段长度50~456 bp。其中,文蛤、青蛤、菲律宾蛤仔和硬壳蛤分别得到681,715,702和694个位点,相应的多态位点比率为76.8%,81.7%,83.0%和75.1%,得到17个种特异性位点,可作为4物种特征标记。分析了群体遗传相似系数和遗传多样性指数以及种间遗传相似系数。结果表明,硬壳蛤群体遗传相似系数最高(0.670 9),遗传多样性指数最低(0.236 0);菲律宾蛤仔群体遗传相似系数最低(0.592 5),遗传多样性指数最高(0.261 8);根据遗传相似系数采用UPGMA法构建了4物种32个体的聚类图,表明文蛤与菲律宾蛤仔遗传关系最近,青蛤与其他3物种遗传关系较远。     

Microbial community structure and diversity in deep-sea hydrothermal vent sediments along the Eastern Lau Spreading Centre

The aim of this study is to investigate microbial structures and diversities in five active hydrothermal fields’ sediments along the Eastern Lau Spreading Centre (ELSC) in the Lau Basin (southwest Pacific). Microbial communities were surveyed by denatured gradient gel electrophoresis (DGGE) and clone library analysis of 16S rRNA genes. The differences in microbial community structures among sediment samples from the five deep-sea hydrothermal sites were revealed by DGGE profiles. Cluster analysis of DGGE profiles separated the five hydrothermal samples into two groups. Four different 16S rRNA gene clone libraries, representing two selected hydrothermal samples (19-4TVG8 and 19-4TVG11), were constructed. Twenty-three and 32 phylotypes were identified from 166 and 160 bacterial clones respectively, including Proteobacteria, Bacteroidetes, Firmicutes, Nitrospirae and Planctomycetes. The phylum Proteobacteria is dominant in both bacterial libraries with a predominance of Gamma-Proteobacteria. A total of 31 and 25 phylotypes were obtained from 160 and 130 archaeal clones respectively, including Miscellaneous Crenarchaeotic Group, Marine Group Ⅰ and Ⅲ, Marine Benthic Group E, Terrestrial Hot Spring Crenarchaeota and Deep-sea Hydrothermal Vent Euryarchaeota. These results show a variety of clones related to those involved in sulfur cycling, suggesting that the cycling and utilization of sulfur compounds may extensively occur in the Lau Basin deep-sea hydrothermal ecosystem.     

长江口泥螺群体遗传多样性的AFLP分析

本研究应用AFLP分子标记技术,对我国长江口泥螺的4个群体(江苏启东,崇明东滩,九段沙湿地国家自然保护区和上海南汇)进行了遗传多样性分析。4对AFLP引物扩增得到577个位点,启东、崇明、九段沙和南汇群体的多态率分别为78.2%、56.0%、58.4%和57.9%。群体总基因多样性为0.2218,显示出较高的遗传多样性水平,且96%以上的遗传变异存在于群体内,群体间的遗传分化极微小(0.0000~0.0404)。群体间Nei’s遗传距离为0.0000~0.0124,主要存在于江苏启东群体和其他3个群体之间。利用STRUCTURE分析群体隐性遗传结构,结果表明长江口泥螺群体有3种遗传来源,同时群体之间有强大的基因流。泥螺较强的扩散能力和海洋开放的环境可能是造成泥螺群体遗传同质性较高的主要原因。     

南海北部陆坡深水区沉积物细菌多样性调查

采用分子生物学手段,调查南海北部陆坡深水区沉积物细菌多样性.分析表明其细菌分成5个类群,变形细菌门(Proteotmcteria),厚壁菌门(Firmicutes),浮霉菌门(Planctomycetes),拟杆菌门(Cytophaga/Flexi-bacteria/Bacteroides,简称CFB)及脱铁杆菌门(Deferribacteres).分别占总体的34%,38%,18%,4%,6%.其中60%以上的克隆子与硫代谢相关,20%克隆子最相近序列来自于油污染环境,说明硫代谢是该区域物质代谢的重要组成,同时该区域有油气渗透的烃类物质存在并影响着该区域的微生物群落结构.     

海南岛东部501站位沉积物古细菌多样性研究

从海南岛东部501站位沉积物中扩增古菌16S rRNA基因构建文库,并对得到的OTU(Operation taxonomic units)进行系统发育学分析.古菌文库由泉古生菌(Crenarchaeota)和广古生菌(Euryarchaeota)构成,其中泉古生菌占总克隆文库的96.8%,由I型海洋泉古生菌(Mrain...     

Phylogenetic analysis of bacterial community in deep-sea sediment from the western Pacific “warm pool”

A depth profile of bacterial community structure in one deep-sea sediment core of the western Pacific ＂warm pool＂ （WP） was investigated and compared with that in a sediment sample from the eastern Pacific （EP） by phylogenetic analysis of 16S rDNA fragments. Five bacterial 16S rDNA clone libraries were constructed, and 133 clones with different restriction fragment length polymorphism （RFLP） patterns were sequenced. A phylogenetic analysis of these sequences revealed that the bacterial diversity in a sample from the WP was more abundant than that in the EP sample. The bacterial population in the sediment core of WP was composed of eight major lineages of the domain bacteria. Among them the γ-Proteobacteria was the predominant and most diverse group in each section of WP sediment core, followed by the α-Proteobacteria. The genus Colwellia belonging to γ-Proteobacteria was predominant in this sample. The shift of bacterial communities among different sections of the WP sediment core was δ-, ε-Proteobacteria, and Cytopahga-Flexibacteria-Bacteroides （CFB） group. The ratios between them in the bacterial communities all showed inversely proportional to the depth of sediment. The sequences related to sulphate reducing bacteria （SRB） were detected in every section. The bacterial community structure in this sediment core might be related to the environmental characteristics of the surface seawater of the western Pacific WP.     

红树植物拟海桑钙调蛋白基因的克隆及分析

钙调蛋白（Calmodulin)是生物细胞内一种重要的调控蛋白，通过其与靶酶的相互作用控制细胞正常的生长和发育，作者以高抗盐红树植物海桑属拟海桑（Sonnera-tia paracasolaris)总DNA为模板，参考GenBank上植物钙调蛋白基因序列合成5′端和3′端引物，利用多聚酶链式反应（PCR）扩增了拟海桑钙调蛋白基因，与克隆载体pBsk( )重组，转化Escherichia coliDH5α得到重组克隆子，DNA序列分析表明，所得片段的编码区在核苷酸序列上与迄今已知的几种植物钙调蛋白基因有很高的同源惺 ，同源率在85%以上；与水稻、苹果，衣藻等相似，其基因编码区被一个位于第75位核苷酸之后的内含子所中断。     

Cloning and Structural Analysis of Calmodulin Gene from the Mangrove Plant Sonneratia Paracaseolaris

Calmodulin is a calcium binding protein that modulates the activity of diverse groups of protein including some protein kinase, adenylate cyclases and ATPase. Here we use the total DNA of Sonneratiaparacaseolaris as the template ofthe polymerase chain reaction (PCR). The PCR primers have been designed and synthesized according to the 5-and 3-terminal oligonucleotide sequences of Calmodulin gene of plants in Genbank and ligated with cloning vector pBsk( ).The recombinant clones have been obtained from the selected medium. The results of DNA sequences analysis show that the nucleotide sequences of ORF share more than 85% homologies as compared with those ofcalmodulin genes of several other plants. Similar to rice and apple, the ORF is interrupted by an intron behind the 75th nucleotide.     

Additions to the echinoid fauna of New Zealand

Seven species of echinoids are added to the New Zealand fauna, two being described as new species of the genera Caenopedina and Spatangus Described species, new to the fauna, are Gracilechinus multidentatus (H. L. Clark), Pourtalesia laguncula A. Agassiz, Hemiaster expergitus gibbosus A. Agassiz, and Gymnopatagus magnus A. Agassiz and H. L. Clark. Fragments of Spatangus, possibly hybrid S. thor Fell, and fragments of Echinocardium, almost certainly not E. cordatum (Pennant), are also described.