运用nrDNA ITS 数据研究角叉菜属(Chondrus)在红藻门中的系统进化地位

测定了角叉菜属(Chondrus)5个代表种的核糖体DNA内转录间隔区(ITS)及5.8SrDNA基因序列。结果表明,角叉菜属ITS区(含ITS1、5.8SrDNA和ITS2)序列长度范围为704—714bp,G+C含量为44.6%—45.7%,变异位点69个,信息位点16个;其中,ITS1和ITS2的长度范围分别为147—149bp和398—404bp。5.8SrDNA长度为158bp,没有变异和信息位点。由MEGA3构建的系统进化树(ME和MP)显示:在进化尺度上,真红藻纲的松节藻科(Rhodomelaceae)与红毛菜纲(Bangiophyceae)亲缘关系较近。在真红藻纲内,杉藻目(Gigartinales)的进化地位相对较高,其次是海膜科(Halymeniaceae)、石花菜科(Gelidiaceae)、红叶藻科(Delesseriaceae)和粉枝藻科(Liagoraceae)等,而松节藻科进化地位相对较低。在杉藻目内,杉藻科(Gigartinaceae)和胶黏藻科(Dumontiaceae)进化关系密切,而形态学特征相似的角叉菜和马泽藻(Mazzaella)亲缘关系非常近。     

广西文蛤(Meretrix)的fAFLP 及ITS 分析

利用fAFLP标记技术和PCR-RFLP技术,对采集于广西的形态和壳色变异很大的2个文蛤群体(G、W)以及山东文蛤(S)群体进行了遗传结构和亲缘关系研究.fAFLP分析显示:在457个总扩增位点中存在53个W群体的特异性位点(其中9个为100%显性)、21个G群体的特异性位点、14个S群体的特异性位点,这些位点可作为群体鉴别的特征性标记;通过计算群体间相对遗传距离和聚类分析,发现S和G间的遗传距离最小(0.0424),在系统树上首先聚在一起,而W与S、G的相对遗传距离均很大(0.2308和0.2305),单独分出一支.对文蛤核糖体ITS的PCR-RFLP结果显示:W的三个不同壳色类群(WX、WC、WH)的酶切结果完全一致,而与G和S群体差别较大;G和S群体基本一致,但也稍有差别.进一步的ITS序列分析显示:W的ITS序列长度为1266-1269bp.而G和S分别为1614bp和1520bp;W群体内部序列比较保守,而与G和S群体之间在ITSl和ITS2开始和结尾处均存在多处插入/缺失;在依据序列构建的系统树上,WX、WC、WH三个类群聚在一起,S和G聚为另一支,两支相距甚远.本研究结果表明,fAFLP标记分析、PCR-RFLP分析和ITS序列分析结果一致,W群体和G、S群体的差异较大,已超出种内群体间的遗传变异,S和G尽管存在遗传分化,但仍应同属Meretrix meretrix-个物种.     

Molecular phylogenetic analysis of attached Ulvaceae species and free-floating Enteromorpha from Qingdao coasts in 2007

Based on the sequence data of the nuclear ribosomal DNA internal transcribed spacer(ITS) 1,5.8 S,and ITS 2,the molecular phylogeny was analyzed on Ulvaceae species collected from Qingdao coasts in summer of 2007,including 15 attached Ulva and Enteromorpha samples from 10 locations and 10 free-floating Enteromorpha samples from seven locations.The result supported the monophyly of all free-floating Enteromorpha samples,implying the unialgal composition of the free-floating Enteromorpha,and the attached Ulvaceae species from Qingdao coasts were grouped into other five clades,suggesting that they were not the biogeographic origin of the free-floating Enteromorpha in that season.     

Molecular phylogenetic analysis of attached Ulvaceae species and free-floating <Emphasis Type="Italic">Enteromorpha</Emphasis> from Qingdao coasts in 2007

Based on the sequence data of the nuclear ribosomal DNA internal transcribed spacer (ITS) 1, 5.8 S, and ITS 2, the molecular phylogeny was analyzed on Ulvaceae species collected from Qingdao coasts in summer of 2007, including 15 attached Ulva and Enteromorpha samples from 10 locations and 10 free-floating Enteromorpha samples from seven locations. The result supported the monophyly of all free-floating Enteromorpha samples, implying the unialgal composition of the free-floating Enteromorpha, and the attached Ulvaceae species from Qingdao coasts were grouped into other five clades, suggesting that they were not the biogeographic origin of the free-floating Enteromorpha in that season. Supported by NSFC (40506030); the Innovative Key Project of the Chinese Academy of Sciences (KZCX2-YW-209); Science & Technology Project of Qingdao City (06-2-2-12-JCH)     

南海海域红斑后海螯虾(Metanephrops thomsoni)的分子识别与群体遗传分化

利用16S rRNA基因序列分子识别红斑后海螯虾,然后对海南附近海域(18°30′-19°00′N,111 °30′-112°30′ E)、北部湾口海域(15°41′ N,110°40′ E)和南沙群岛海域(5°20′-5°29′N,110°09′-111°26′E)三个地理群体进行ITS1的扩增测序.16S rRNA基因序列的结果表明所取样本准确;三个地理群体分别得到616-623bp、619bp和614bp的ITS1全长序列,A、G、T、C含量平均分别为22.3％、29.4％、17.8％和30.6％;其中562个保守位点,39个多态位点.ML和NJ聚类分析显示三个地理群体共聚为三个大支,其中海南群体有群体分化.表明红斑后海螯虾三个地理群体分化极其显著,且海南群体具有相对高的遗传多样性.     

东海三叶原甲藻的形态特征及其ITS序列分析

报道了首次分离于东海海域的三叶原甲藻(Prorocentrum triestinum Schiller)藻株号(LAMB100721),通过利用光学显微镜、荧光显微镜、扫描电镜及分子生物学方法,对其形态特征、显微结构和分子系统进化进行了详尽描述和鉴定。细胞长卵形或披针形,后端细长且尖,前端圆,最宽部位于细胞中央。顶刺长而显,三角状。壳面光滑,无刺或突起物。刺丝胞孔稀疏而不规则地分布于壳面边缘。叶绿体无具体形状,分布于整个细胞中,细胞核球形,位于中下部。老化的细胞可见细胞边缘的间接带,且间接带表面光滑。细胞长为19~25 μm,平均值为(22.6±0.9)μm;宽为11~16 μm,平均值为(13.2±1.1) μm。所测目标藻株的rDNA ITS序列长度为569 bp,其中GC含量为47.6%。三叶原甲藻系赤潮种,为东海原甲藻春季大规模赤潮的伴随种。加强有害赤潮的预防和监测工作是减少危害的有效途径,而对赤潮原因种的准确识别和鉴定则是基础和关键。     

Complete nuclear ribosomal DNA sequence analyses of the black-footed abalone Haliotis iris

The black-footed abalone Haliotis iris is an economically important shellfish species in New Zealand. We successfully amplified, sequenced and analysed the complete nuclear ribosomal DNA (nrDNA) of H. iris. The length of the nrDNA was determined to be around 9.6?kb and included, in order, small subunit ribosomal RNA (nrSSU, 1858bp), internal transcribed spacer (ITS, 749?bp), large subunit ribosomal RNA (nrLSU, 3412bp) and an intergenic spacer (IGS, 3560–3662?bp). The nrLSU genes were identical in two individuals, whereas the nrSSU and ITS regions existed at three and four base differences, respectively. The IGS was more variable than the other nrDNA regions. A phylogenetic tree was constructed based on the ITS sequence datasets, which revealed that Haliotidae has two major subclades, mainly distributed in the North Pacific, Europe and Australia. The complete nrDNA sequence will be useful for the classification, phylogeny and breeding of this shellfish.     

Sensitive and rapid detection of two toxic microalgae Alexandrium by loop-mediated isothermal amplification

A loop-mediated isothermal amplification (LAMP) assay was designed and evaluated for rapid detection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning (PSP). Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer (ITS) of ribosomal DNA. The method worked well in less than an hour under isothermal conditions of 65℃. LAMP specificity was validated in closely related algae as a comparison, suggesting the strict specificity of the LAMP primers. Two visual inspection approaches were feasible to interpret the positive or negative results. The detection limits of A. catenella and A. minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA, respectively. The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction (PCR) method in detecting the two microalgae. These characteristics of species specificity, sensitivity, and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A. catenella and A. minutum.     

发现于我国渤海辽东湾的海洋甲藻新种-渤海异冒藻(Heterocapsa bohaiensis sp. nov.)

A small armed dinoflagellate bloomed in the aquaculture ponds off the coast of Liaodong Bay, Bohai Sea of China,resulting in heavy mortalities of the cultured prawns(Penaeus japonicus) and larvae of Chinese mitten handed crabs(Eriocheir sinensis). The bloom-forming species was successfully isolated, and cellular morphology of the specimen was consequently investigated through light, fluorescent and electron microscopy. The small((14.4±1.6) μm in length) ellipsoid cells show typical Heterocapsa thecal plate arrangement(Po, cp, 5′, 3 a, 7′′, 6 c,5 s, 5′′′, 2′′′′). The episome is evidently bigger than the hyposome. One to three spherical pyrenoids are located above or beside the large elongated nucleus. The body scale is characterized by a triangle basal plate with one central upright and nine peripheral spines. Above all, Heterocapsa bohaiensis could be distinguished from other Heterocapsa species by the combination of the cell size, morphology, cellular structure and body scale. Sequence analyses of both ITS and LSU regions reveal the significant genetic divergence between H. bohaiensis and other established species in this genus, further supporting novelty of this species. Noticeably, different sample treatment methods resulted in morphological variation of the apical pore complex(APC) of H. bohaiensis, which needs to be taken into account in future study.     

Palaeoenvironmental interpretation of Holocene coastal sequences in the southern bay of Biscay

The main sedimentary features of the northern coast of the Iberian Peninsula during the Holocene transgression are characterized by the formation of estuaries, the deposition of sand bars and sand beaches, and the accumulation of aeolian dunes. These coastal deposits are very favourable for identifying Quaternary sea-level changes as they contain great volumes of well preserved sediments including marine, brackish and freshwater beds. The micropalaeontological analysis (benthic foraminifera) of diverse littoral sequences has allowed different microfaunal assemblages to be recognised and their corresponding depositional environments determined. Two recurrent phases of sediment build-up as the sea level rose have been distinguished. They have been interpreted as the consequence of two different marine advances in this region: one dated at around 8 000 years BP and the second around 2 500 years BP.