笛鲷属(Lutjanus)鱼类线粒体16S rRNA基因序列比较及系统学分析

对笛鲷属9种鱼的线粒体DNA16S rRNA基因片段进行了PCR扩增,扩增产物经纯化后测序,得到了长度为561bp的分析序列。结合GenBank中的另外9种笛鲷的16S rRNA同源序列计算得出A、T、G、C的含量平均为28.5%、22.1%、23.6%和25.8%,AT含量稍高于GC含量,18种笛鲷碱基组成差异不大。利用MEGA3.1软件对所得序列进行比对后检测到72个变异位点,其中包括简约信息位点44个,在变异位点中75.61%的碱基替换是由于发生了转换,转换/颠换平均为3.1︰1。计算了种间的遗传距离,结果表明,序列差异在0.0193-0.0680之间,其中勒氏笛鲷和金焰笛鲷、勒氏笛鲷和金带笛鲷的序列差异最小,而红鳍笛鲷和金焰笛鲷、红鳍笛鲷和画眉笛鲷的序列差异最大。选用高体四长棘鲷(Argyrops spinifer)为外群,利用NJ法构建了分子系统树,结果表明:本研究中的9种南海笛鲷与其它9种笛鲷进化关系较远;并且南海9种笛鲷相互之间仍然保持着着相对较远的遗传距离,遗传多样性较好。     

相手蟹属两种蟹类线粒体16SrRNA基因序列的比较

测定了相手蟹属（Sesarma）红螯相手蟹（S．haematocheir）和褶痕相手蟹（S．plicata）线粒体16SrRNA基因部分片段的序列,二者的序列长度相同,均为533bp,且A、T、G、C的含量相似,分别为198bp（37．1％）,206bp（38．6％）,84bp（15．8％）,45bp（8．4％）和200bp（37．5％）,205bp（38．5％）,81bp（15．2％）,47bp（8．8％）;二者的序列有49处差异,其中21个位点为转换、22个位点为颠换和6个缺失／插入位点。进一步对20种相手蟹属蟹类的长度为361bp的16SrRNA基因同源序列进行分析,发现AT的含量为78．6％～82．9％,明显高于GC的含量,且存有91个变异位点。从NJ树和遗传距离来看,在分布于中国的3种相手蟹中,无齿相手型（S．dehaani）和红螯相手蟹的亲缘关系最近（d=0．0151）,而它们与褶痕相手蟹的亲缘关系则较远（d=0．0924／0．09231。分布于中国的相手蟹和分布于北美的相手蟹之间存在着较大的遗传距离（差异）,表明它们之间有着较远的亲缘关系,互为单系起源。     

The divergent environmental characteristics of permanently-open,seasonally-open and normally-closed estuaries of south-western Australia

This study has compared the environmental characteristics of the basins and saline lower reaches of the tributaries of eight estuaries on the south coast of Western Australia, which differ in their degree of connectivity with the ocean. Although four estuaries between 115.1° and 121.8° E along that coast remain permanently open to the ocean, the others only become open when the volume of river discharge is sufficient to breach the prominent sand bars at their mouths, which occurs annually following heavy winter and early spring rainfall in some estuaries (seasonally open) and infrequently in others (normally closed). Estuaries to the west of 118.5° E are predominantly permanently open, e.g. Oyster Harbour, or seasonally open, e.g. Broke, Irwin and Wilson inlets, whereas those further east, e.g. Wellstead Estuary and Hamersley, Culham and Stokes inlets, where annual rainfall and thus discharge are much lower, only become open after exceptionally heavy discharge. In permanently and seasonally-open estuaries, pronounced haloclines and oxyclines are present in the narrow rivers but not the wide basins where the waters are subjected to wind-driven mixing. The extent of cyclical seasonal fluctuations in environmental conditions differed markedly among the three seasonally-open estuaries and between years in one of those systems. These differences reflected variations in the relationship between the volume of fluvial discharge, which is determined by a combination of the amount of local rainfall, catchment size and extent of clearing of native vegetation, and the amount of intrusion by marine waters, which is largely controlled by the size and duration of the opening of the estuary mouth. The mean seasonal salinities in the basins of the three normally-closed estuaries increased over three years of very low rainfall to 64 in the deepest of these estuaries (Stokes Inlet) to 145 in Hamersley Inlet and to 296 in the shallowest estuary (Culham Inlet). These results demonstrate that the environmental characteristics of estuaries on the south coast of Western Australia differ markedly, even among those of the same type, e.g. seasonally-open estuaries or normally-closed estuaries.     

Profiling Planctomycetales diversity with reference to anammox‐related bacteria in a South China Sea,deep‐sea sediment

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.     

黄海西北近岸沉积物中细菌群落空间分布特征

采用16S rDNA文库和变性梯度凝胶电泳(DGGE)技术,对黄海西北部3个近岸站位沉积物中细菌群落多样性及空间分布特征进行了调查和解析。对表层沉积物16S rDNA序列统计表明,各站位细菌群落多样性很高,γ-和δ-变形菌纲分别占克隆序列总数的20%~32%,是沉积物中的绝对优势类群。DGGE图谱分析表明,同一站位中不同深度的细菌群落结构相似性较高,而不同站位间群落结构相差较远。研究表明在黄海西北近岸沉积物中细菌群落多样性较高,优势类群明显,在较小尺度范围内群落结构的垂直变化不明显。     

冬季刺参养殖环境与肠道内细菌菌群的研究

运用传统细菌分离培养与分子生物学技术相结合的方法,对2008年11月至2009年1月冬季刺参(Apostichopus japonicus Liao)养殖池塘环境(养殖水、底泥、附着基)及刺参肠道内的细菌菌群进行了分析。应用平板稀释涂布培养计数法测得刺参养殖池塘水体、底泥、附着基和肠道细菌数量分别为0.75×102～1.4×104cfu/mL、8.7×104～8.1×105cfu/g、3.8×105～2.8×106cfu/g、7.1×105～1.5×107cfu/g;根据形态学差异从培养所得的细菌中筛选得到22株菌,用限制性内切酶Rsa I和Msp I对所分离菌株进行ARDRA(Amplified rDNA Restriction Analysis)分析,这22株菌被分为8种不同的分类单元(Operational Taxonomic Unit,OTU),其中OTU2与OTU3所包含的菌株分别占分离菌株种数的30%和20%;此外,作者对不同环境培养所得的优势度最高的细菌进行分子鉴定分析。结果表明:水环境中优势菌为施氏假单胞菌(Pseudomonas stutzeri)、门多萨假单胞菌(Pseudomonas mendocina)及巨大芽孢杆菌(Bacillus megaterium),沉积物中优势菌为巨大芽孢杆菌(B.megaterium)、施氏假单胞菌(P.stutzeri)、苏云金芽孢杆菌(Bacillus thuringiensis),附着基中优势菌为巨大芽孢杆菌(B.megaterium)、苏云金芽孢杆菌(B.thuringiensis),灿烂弧菌(Vibrio splendidus),肠道中优势菌为巨大芽孢杆菌(B.megaterium)、苏云金芽孢杆菌(B.thuringiensis),灿烂弧菌(V.splendidus)、施氏假单胞菌(P.stutzeri)。通过对冬季刺参池塘细菌菌群多样性分析和优势菌鉴定,为筛选低温益生菌和防治刺参疾病提供了有益参考。     

中国沿海10种方蟹16S rRNA基因序列分析及系统发育研究

中国沿海10种方蟹线粒体16S rRNA基因部分片段的序列长度为517 bp～533 bp。它们的核苷酸序列A、T、G、C的含量相似,A+T的含量(69.8%～76.0%)明显高于G+C的含量;10种方蟹的16S rRNA基因序列比对获得541 bp的同源序列(含插入/缺失位点),除插入/缺失位点外共检测到146个变异位点,其中81个为简约信息位点。4种厚蟹与2种近方蟹的遗传距离(0.054～0.085)都显著小于与其他方蟹之间的遗传距离,甚至明显小于与4种厚蟹原本属于同一相手蟹科的2种相手蟹之间的遗传距离(0.105～0.155);而基于16S rRNA基因片段序列采用NJ法构建的系统进化树的拓扑结构也显示,原本属于相手蟹科的侧足厚蟹、天津厚蟹、日本仿厚蟹和伍氏仿厚蟹没有与2种相手蟹聚为一支,而是最终与属于弓蟹科的2种近方蟹聚为一大支,且有高达99%的支持率。结果表明,4种厚蟹与2种近方蟹的亲缘关系相对较近,而与2种相手蟹等其他方蟹的亲缘关系则相对较远。因此,研究结果支持将4种厚蟹从相手蟹科移到弓蟹科。此外,属于相手蟹科的2种相手蟹聚为一支,属于方蟹科的白纹方蟹和属于斜纹蟹科的瘤突斜纹蟹又各自成为一支;表明16S rRNA基因的分子数据支持其形态学分类结果的正确性,提示上述4科蟹类可能分别为单系。     

两种体色三疣梭子蟹线粒体DNA 部分片段序列的比较分析

三疣梭子蟹(Portunus trituberculatus)作为重要的海洋经济动物,其常见体色(头胸甲)为茶绿色。近年来在我国沿海海域海捕三疣梭子蟹中开始出现体色为紫色的一类梭子蟹,除体色不同外,二者表型特征并无显著差异。对两种体色三疣梭子蟹不同个体的线粒体COI和16S rRNA基因进行了序列比对分析以判定二者之间的亲缘关系。结果发现COI和16S rRNA基因在两种体色三疣梭子蟹群体中的核苷酸序列一致性分别为99.87%和99.88%,表明紫色三疣梭子蟹并未发生亚种的分化,即紫色和茶绿色群体属于同一个种。并利用GenBank数据库检索了梭子蟹科其他15种海产蟹的16S rRNA序列,进行了序列同源性比对分析和分子系统学方面的探讨,为梭子蟹种属分类学研究提供分子依据。     

Comparative research on karyotype in three species of Arcidae

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Microbial Diversity in Nankai Trough Sediments at a Depth of 3,843 m

Dense populations of bivalves, primarily Calyptogena sp., were observed at cold seeps of the Nankai Trough. Bacterial input to the sediment was estimated through determination of phospholipid ester-linked fatty acid (PLFA) and DNA profiles. Results indicated a bacterial biomass of 10⁹ cells (g dry wt)^-1 while individual fatty acid profiles revealed a predominance of monounsaturated fatty acids, mainly 18:1 isomers. The presence of these fatty acids can be interpreted to reflect a response to low temperature and a predominance of psychrophilic bacteria. DNA fragments encoding bacterial ribosomal RNA small-subunit sequences (16S rDNA) were amplified by the polymerase chain reaction method using DNA extracted directly from the sediment samples. From the sequencing results, at least 19 kinds of bacterial 16S rDNAs related to mostly the Proteobacteria and a few gram-positive bacteria were identified. These results suggest that the bacterial community in the Nankai Trough sediments consists of mainly bacteria belonging to the Proteobacteria , , and subdivisions. Bacteria belonging to the and subdivisions, which are known to include epibiont and sulfate reducing bacteria, respectively, were mostly detected in the sediment obtained from inside the area of the Calyptogena community, and the -Proteobacteria may function to supply reduced sulfur to bacterial endosymbionts of Calyptogena.