首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到18条相似文献,搜索用时 109 毫秒
1.
东太平洋深海沉积物中DNA的提取及细菌多样性初步分析   总被引:1,自引:0,他引:1  
以东太平洋海隆附近深海柱状沉积物为材料,通过化学裂解和酶消化相结合的方法提取了沉积物微生物的总基因组DNA,并进行了纯化。结果表明所得到的DNA分子片段大小在21kb左右,纯化后的DNA可直接用于PCR等分子生物学操作。细菌16SrDNAV3可变区的PCR—DGGE图谱展示出15条以上条带,表明深海沉积物中细菌多样性较高,群落结构比较复杂。对其中9条主要条带进行回收、测序和系统发育分析,结果表明所获得的序列分属放线菌门(Actinobacteria),绿弯菌门(Chloroflexi),γ-变形细菌亚门(Gamma—proteobacteria),α-变形细菌亚门(Alpha—proteobacteria)和嗜酸菌门(Acidobacteria)5个大类群。  相似文献   

2.
东太平洋海隆深海热液区沉积物微生物多样性的研究   总被引:2,自引:1,他引:1  
提取东太平洋海隆区深海热液系统沉积物样品的总DNA,构建沉积物中的细菌16S rDNA 克隆文库,通过PCR-RFLP分析与序列测定,对沉积物中的微生物类群及其与环境的关系进行了分析.结果表明,该海区沉积物中的36个克隆代表的22种基因型分别属于7个主要类群,其中变形菌(Proteobacteria)的γ-亚群为优势菌群,α-和β-亚群也均有分布;而硫氧化相关共生菌的属(sulfur-oxidizing symbionts)为优势种属.系统发育分析表明,在该沉积物中细菌主要是跟共生有关、跟C、S代谢相关,大多还能在无氧和高温环境的条件下生存,说明采样点具有典型的深海热液生态系统的特点,甲烷代谢和硫代谢在该区域的深海物质能量循环中占据着重要地位.另外大量新的极端微生物的存在,预示着该区域的微生物资源有着非常大的开发潜力.  相似文献   

3.
沉积物成分复杂,腐殖酸等物质含量较高,这些杂质在DNA提取过程中不易除去,可能会对后续的PCR扩增等产生抑制作用。因此,高质量DNA的获得是海洋微生物分子生态学研究的基础和前提。本研究采用6种不同的方法,分别提取同一来源海洋沉积物样品中的微生物基因组DNA。对DNA纯度、得率、16SrRNA基因拷贝数和微生物群落特征等多方面进行比较,结果表明:方法1(QIAamp DNA Stool Mini Kit)、方法2(PowerSoil~DNA Isolation Kit)和方法3(RNA PowerSoil~DNA Elution Accessory Kit)得到的DNA产量较低,纯度却较高,可以直接用于后续分子生物学分析;与方法2相比,方法3得到的细菌16SrRNA基因拷贝数和单位质量DNA中可扩增细菌16SrRNA基因模板量较低;方法4(SDS高盐法)得到的DNA虽然产量高,但杂质含量也高,抑制了后续PCR反应的进行;方法5(方法4得到的DNA经QIAamp DNA Stool Mini Kit纯化)和方法6(方法4得到的DNA经PowerSoil~DNA Isolation Kit纯化),虽然纯度有所提高,但DNA大量损失,且耗时长,花费高。另外,方法2在提取过程中可以最大程度裂解菌体细胞壁,能更好反映微生物群落特征。综合以上结果,方法2(PowerSoil~DNA Isolation Kit)提取海洋沉积物微生物基因组DNA效果最佳。本研究可为海洋微生物分子生态学研究提供一种有效的技术手段。  相似文献   

4.
北极海洋沉积物中可培养细菌及其多样性分析   总被引:2,自引:1,他引:1  
王桢  李阳  车帅  林学政 《海洋学报》2014,36(10):116-123
利用Zobell 2216E培养基和涂布平板法对北极海洋沉积物中可培养细菌进行分离纯化,并利用16SrRNA基因进行分子鉴定与系统发育分析。根据菌落形态学特征,从59个站点的沉积物样品中共分离纯化获得570株细菌;基于16SrRNA基因的分子鉴定与系统发育分析表明,分离到的可培养细菌分别属于细菌域的4个门,5个纲,12个目,23个科,47个属,102个种,其中γ-Proteobactria占绝大多数;有14株菌株与模式菌株的16SrRNA基因序列相似性小于97%,为6个潜在的新种。北极海域的海洋沉积物中存在着丰富的微生物种质资源,为开发新型生物活性物质和特殊功能基因打下了基础。  相似文献   

5.
采用化学裂解法从乳山湾外海溶氧低值区不同溶解氧质量浓度(3.7~7.0mg.L-1)的6个站位海水样品中提取了环境DNA样品。以试剂盒纯化后的DNA样品为模板扩增其16SrRNA基因V3区,通过变性梯度凝胶电泳、分子文库构建及DNA测序对溶氧低值区海水中的细菌群落结构进行研究。结果表明,乳山湾外海溶氧低值区不同溶解氧质量浓度的6个站位底层海水样品中的细菌群落结构是相似的,它们均由隶属于Alteromonas(交替单胞菌属)、Salegentibacter(需盐杆菌属)等10个属的18种细菌组成。系统发育分析发现这些细菌分别属于α变形菌纲(2种)、γ变形菌纲(12种)和黄杆菌纲(3种)三个大类。在乳山湾外海溶氧低值区的海水样品中细菌多样性最高的类群是γ变形菌纲。  相似文献   

6.
南海北部巴士海峡深海沉积物中细菌多样性分析   总被引:2,自引:1,他引:1       下载免费PDF全文
从南海北部巴士海峡深海沉积物中提取到高质量的总DNA, 通过TA克隆构建了含有23个可操作分类单元(OTU)的16S rRNA基因文库, 选择各OTU中代表性克隆子进行测序与系统发育分析, 结果表明, 南海北部巴士海峡深海沉积物中细菌在系统进化树中至少分属于9个类群: 其中放线菌Actinobacteria, 变形细菌Proteobacteria, 和浮霉菌Planctomycetes为优势种;其他细菌如疣微菌Verrucomicrobia, 鞘脂杆菌Sphingobacteria, 硝化螺旋菌门(Nitrospira), 绿弯菌Chloroflexi, 厚壁菌Firmicute, 酸杆菌Acidobacteria等为非优势种群;另外有两个克隆子属于未知种群.在所获得的23个代表克隆子序列中,有11个序列与已知细菌的同源性≤95%, 占到了所有序列的48%, 这一结果说明在南海北部巴士海峡海区具有非常丰富的微生物多样性, 这一海区蕴含着大量未知的微生物资源, 因而值得进行进一步的研究探索.  相似文献   

7.
北极深海沉积物中细菌和古菌群落结构研究   总被引:3,自引:0,他引:3  
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.  相似文献   

8.
提取西太平洋"暖池"区深海沉积物样品总DNA,利用异化型亚硫酸盐还原酶(DSR)和甲基辅酶M还原酶(MCR)基因的特异引物,采用PCR-RFLP方法对沉积物样品中这两类功能基因多样性进行研究.该沉积物中的dsrAB基因分别来源于δ-变形菌中的6个属,其中最多的是脱硫弧菌属和脱硫杆状菌属;mcrA基因均来源于产甲烷古菌,其中主要是甲烷微菌.这些基因在系统发育树上都处于相对独立的分支.此外,还有较多的dsrAB,mcrA基因来源于未知的新属或新种.这些结果表明该海区沉积物中由微生物参与的硫和甲烷循环比较活跃,而且其中可能存在多种新的代谢途径.  相似文献   

9.
环境DNA(eDNA)技术结合高通量测序已广泛应用于评价微型生物多样性与群落构成。相较于eDNA,RNA在环境中易降解,环境RNA(eRNA)可更准确地反映群落近期生命活性状态。理论上,基于eRNA检获的生物多样性要低于eDNA检获的总体多样性。但前期已发表研究显示同一站点eDNA获得的多样性低于eRNA检获量,推测可能原因包括:1)样本量不同;2)RNA中存在DNA污染。为验证假设,本研究以陆架区两个站位沉积物中的纤毛虫为实验对象,系统性比较四种DNA/RNA提取方法:DNA直接提取(0.9g沉积物),大样本量DNA直接提取(2g),DNA洗脱法(2g),RNA直接提取法(2g);以及三种RNA提取后的处理方法:无DNA酶反转录,含DNA酶反转录,先纯化RNA再反转录。研究结果表明,大样本量(2g)DNA直接提取法所检获的可操作分类单元(OTUs)约为小样本量(0.9g)的两倍。相同样本量下,DNA洗脱法检获的OTUs最多,而DNA直接提取检获的OTUs低于有/无DNA酶反转检获的数量,先纯化再反转录所获得OTUs最少。就群落构成而言,DNA洗脱法和RNA纯化可有效降低浮游类群比例,可更真实展现底栖群落的构成。综上,建议使用DNA洗脱法评价沉积物中微型生物的总体多样性;采用eRNA研究具有生命活性的生物群落时,反转录之前可纯化RNA,以获得更加准确的具有生命活性的群落信息。  相似文献   

10.
南海沉积物中有孔虫分子生物学鉴定的初步研究   总被引:1,自引:0,他引:1  
徐美香  赵泉鸿  肖湘 《台湾海峡》2004,23(3):354-359
本文以南海深海表层沉积物中的有孔虫样品为研究对象,提取了样品的总DNA片段,利用特异性DNA探针,进行有孔虫的SSUrDNA扩增,对获得的PCR产物进行克隆,测序及分析,证实确为潜在的有孔虫DNA.进化树分析揭示了研究样品中可能存在的有孔虫种类,该方法的建立为有孔虫壳体的鉴定提供了新的技术手段  相似文献   

11.
12.
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.  相似文献   

13.
Although environmental factors such as grain size and organic carbon content may influence the distribution of microbes in marine sediments, there has been little experimental study of the topic to date. To investigate how those sediment variables affect microbial colonisation under in situ conditions, deep‐sea sediments and artificial sediments (glass beads, sands) were incubated in the Arctic deep sea at 2500 m water depth with or without chitin, one of the most important carbon polymers in marine environments. Microbial abundance, biomass, chitobiase activity and changes in community structure were monitored after 7 days and 1 year. In control sediments without chitin addition, no significant changes in microbial abundance, biomass and activity were observed after 1 year. In the presence of chitin, however, considerable increases in these parameters were recorded in all three sediment types tested. Regardless of chitin addition, natural deep‐sea sediments were always associated with higher values of microbial abundance, biomass and activity compared with artificial sediments. Sediment type was always found to be the most significant factor explaining variation in enzymatic activity and bacterial community structure as compared to the effects of chitin amount, incubation time, and changes in cell number or biomass. Overall, this is the first in situ study that has addressed the effects of multiple factors and their interactions on abundance, biomass, activity and community structure of microbial communities in the deep Arctic Ocean.  相似文献   

14.
将16S rDNA技术应用于近海环境监测中,分析海洋环境样品中的微生物组成与环境污染程度的关系.从原核微生物多样性的改变较为准确地确定海洋污染程度,是传统海洋环境监测的良好补充,有利于加大近海海洋环境监测力度。  相似文献   

15.
东海陆架表层沉积物微生物多样性初步研究   总被引:2,自引:2,他引:0  
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。  相似文献   

16.
A systematic assessment of Planctomycetales diversity in a South China Sea, deep‐sea sediment (1657 m) was conducted using the 16S rRNA gene analysis approach. PCR amplification of the samples from seven sediment layers (0.1, 1, 3, 5, 7, 9 and 11 m below the surface sediment) using the primer set Pla‐46‐F/1392‐R showed that the Planctomycetales existed within a limited range of sediment depths (≤ 5 m), and had a decreasing trend in diversity with increasing depth. The majority of the retrieved Pla‐46‐F/1392‐R sequences belonged to Pirellula‐related Planctomycetales, and two sequences retrieved from the 0.1‐m layer (GenBank accession numbers: DQ996944 and DQ996945 ) shared the same anammox‐related signature oligonucleotides and were closely related to commonly recognized anammox organisms. To identify new anammox‐related biomarkers, three primer sets were designed for amplifying the fragments of hydroxylamine oxidoreductase and S‐adenosylmethionine radical enzyme genes, but no related sequences were found. Our multiple 16S rRNA gene primer sets (Journal of Rapid Methods and Automation in Microbiology, 2008, in press) revealed even an higher diversity of Planctomycetales in the 0.1‐m layer of the sediment, especially at genus level. Our data profiled the distribution pattern of Planctomycetales diversity along sediment depths, and provided molecular evidence for the existence of anammox‐related bacteria in a new location, which broadens our understanding of Planctomycetales diversity in deep sea sediments.  相似文献   

17.
The bacterial diversity in surface sediment from the South China Sea   总被引:1,自引:0,他引:1  
16S rDNA sequencing results from this study and literatures demonstrate that sediment bacteria in the South China Sea (SCS) were very diverse,which contained 22 of the 24 phyla of bacteria investigated from marine sediment,however,it was very imbalance among stations.So bacterial diversity from 15 samples which covered a wide range of sediment types from 20 to 3 888 m in depth was studied in DGGE (denature gradient gel electrophoresis) in this paper.The DGGE results indicate that both sediment bacterial diversity and diversity difference among stations were significant.Thirty representative and differential fingerprints among samples were recovered and sequenced,phylogenetic analysis indicates that they may belong to Proteobacteria (α-,β-,γ-,δ-,ε-),Planctomycetes,Firmicutes,Chloroflexi,Acidobacteria,Actinobacteria,Nitrospirae,Gemmatimonadetes,candidate division WS3 and so on,of which,Gemmatimonadetes and candidate division WS3 bacteria were first detected in SCS sediment.This study also shows that bacterial diversity analysis based on DGGE was more potential than traditional 16S rDNA clone library in multiple sample analysis.  相似文献   

18.
Studying the diversity‐ecosystem function relationship in the deep sea is of primary importance in the face of biodiversity loss and for our understanding of how the deep sea functions. Results from the first study of diversity‐ecosystem function relationships in the deep sea (Danovaro et al. 2008; Current Biology, 18, 1–8) are unexpected and show an exponential relationship between deep‐sea nematode diversity and ecosystem function and efficiency, although this relationship appears largely restricted to relatively low diversities [ES(51) <25]. Here, we investigate the relationship between nematode diversity and several independent measures/proxies of ecosystem function (sediment community oxygen consumption, bacterial biomass, bacterial extracellular enzyme activity) and efficiency (ratio of bacterial/nematode carbon to organic C content of the sediment) on the New Zealand continental slope. Nematode diversity at our study sites was relatively high [ES(51) = 30–42], and there was no relationship between species/functional diversity and ecosystem function/efficiency after accounting for the effects of water depth and food availability. Our results are consistent with a breakdown of the exponential diversity‐function relationship at high levels of diversity, which may be due to increased competition or greater functional redundancy. Future studies need to take into account as many environmental factors and as wide a range of diversities as possible to provide further insights into the diversity‐ecosystem function relationship in the largest ecosystem on Earth.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号