蛤蜊科3种贝类16SrRNA基因片段及ITS2核苷酸序列分析

利用PCR技术分别扩增连云港及启东沿海蛤蜊科的西施舌（Coelomactra antiquata）、中国蛤蜊（Mactra chinensis）和四角蛤蜊（Mactra veneriformis）3种双壳贝的16SrRNA基因片段和ITS2核苷酸序列．测序后用DNAstar软件分析了核苷酸差异。结果显示：三种贝类16SrRNA基因片段长度相同，均为306bp（去除引物）．核苷酸存在多态性。共有45个变异位点，54个核苷酸发生了变异。全部为碱基置换。西施舌与中国蛤蜊此片段核苷酸的同源性为88．9％．与四角蛤蜊的同源性为88．6％．中国蛤蜊与四角蛤蜊的同源性为90．6％。三种蛤蜊ITS2序列分别为390bp（西施舌）、441bp（四角蛤蜊）和466bp（中国蛤蜊）。存在长度多态性．ITS2核苷酸差异分析结果显示．西施舌与中国蛤蜊的同源性为70．9％-71．1％，西施舌与四角蛤蜊的为70．5％-71．0％。中国蛤蜊与四角蛤蜊的同源性为88．1％-88．8％。ITS2序列分析结果与16SrRNA基因片段分析结果一致．2种分子分析法均显示中国蛤蜊与四角蛤蜊的亲缘关系近。     

Molecular identification of bloom-forming species Phaeocystis globosa (Pryninesiophyta) and its dispersal based on rDNA ITS sequence analysis

The colony-forming Phaeocystis species are causative agents of dense bloom occurrences in coastal waters worldwide. It is difficult to separate them because of the different morphologies associated with their colonial stages. In this study we applied molecular approaches to analyze the genetic variation of Phaeocystis globosa and Phaeocystis pouchetii from several geographic regions, and to assist in tracing the dispersal of bloom-forming Phaeocystis species in coastal waters of China. The sequences of the internal transcribed spacers (ITS1 and ITS2) of rDNA and the 5.8S ribosomal RNA gene of Phaeocystis strains were determined. Sequence comparison shows that P.globosa was the most divergent to P. pouchetii, exhibiting sequence divergence higher than 0.08. However, lower genetic divergences existed between strains of P.globosa. The sequence comparison of the Phaeocystis rDNA ITS clearly shows that the species isolated from the southeast coast of China is identified as P.globosa rather than P. cf. pouchetii or other species. Furthermore, the significance of rDNA variation in distinct global populations of P.globosa suggested it might have had sufficient time to accumulate detectable mutations at the rDNA locus, supporting the hypothesis of ancient dispersal of P.globosa to many areas, meaning that P.globosa blooms in the coastal waters of China are endemic rather than a newly introduced species or a foreign source. Finally, based on the high divergent region of rDNA ITS, a pair of species-specific primers for P.globosa were designed, they could be useful to detect the presence of this species in mixed plankton assemblages or flagellate stages that are recognized with diffculties by means of conventional microscopy.     

Molecular identification of bloom-forming species Phaeocystis globosa(Prymnesiophyta)and its dispersal based on rDNA ITS sequence analysis

Abstract-The colony-forming Phaeocystis species are causative agents of dense bloom occurrences incoastal waters worldwide. It is difficult to separate them because of the different morphologies associatedwith their colonial stages. In this study we applied molecular approaches to analyze the genetic variationof Phaeocystis globosa and Phaeocystis pouchetii from several geogrtaphic regions, and to assist in tracingthe dispersal of bloom-forming Phaeocystis species in coastal waters of China. The Sequences of the inter-nal transcribed sacers (ITS1 and ITS2) of rDNA and the 5.8S ribosomal RNA gene of Phaeocystisstrains were determined. Sequence comparison shows that P.globosa was the most divergent toP. pouchetii, exhibiting sequence divergence higher than 0.08. However, lower genetic divergences ex-isted between strains of P.globosa. The sequence comparison of the Phaeocystis rDNA ITS clearlyshows that the species isolated from the southeast coast of China is identified as P. globosa rather thanP. cf. po     

赤潮叉角藻18SrDNA和ITS区序列测定与分析

采用PCR及克隆测序的方法，对1998年引发渤海赤潮的叉角藻18SrRNAadldey DNAITS区（Internal Transcribed Spacer Regions)进行了序列测定与分析。并通过因特网从国际分子生物学数据库中获取甲藻另外15个种的18rDNA序列，以Tetrahymena corlissi作为类群，分别采用Neighbor-Joining和Fitch方法构建甲藻较为一致和可靠的进化树图，探讨具有高度多样性和在分类上争议较多的甲藻各类群之间的形态与分子进化关系。结果表明，Prorocentrum(有2个简单的壳板）出现得较早，而大多数多甲藻目（覆盖着多个壳板）、裸甲藻目（大多数不具壳板）和膝沟藻目的成员较晚出现。另外，对叉角藻ITS区的分析表明，ITS区为高变区，是良好的分子标记，可用于叉角藻快速鉴定的专一性核酸分子探针的研制。     

智能交通系统中的公交车辆指挥调度体系结构

该文阐述公交智能指挥调度系统总体框架，结合智能交通系统ITS(Intelligent Transport Systems)对公交智能化调度的逻辑结构和物理结构要求，分析公交车辆智能化调度系统结构体系及其模型化，探讨智能调度策略和调度机理。根据该系统总体设计框架构思，了解系统各功能间的相互关系，优化公交车辆运营管理模式，以改善现有公交系统的管理水平以及运营效率，为公交指挥调度系统的完善、提高提供可靠技术基础。     

睡莲类植物ITS nrDNA序列的分子系统发育分析

睡莲类植物是目前植物分子系统学与进化研究的一个重要类群。核基因组的ITS区是核核糖体（nrDNA)转录单位的一部分。测定和分析了5个属的7种睡莲类植物的核核糖体ITS序列，并与GenBank中提取的相关序列进行了组合分析，初步获得了睡莲类植物的2个ITS系统树，结果也支持现有分子系统学研究中有关金鱼藻处于较原始位置的观点。     

GIS的决策支持作用及其在ITS中的应用

地理信息系统(GIS)在智能交通系统(ITS)服务的各个领域中的框架定位作用越来越重要。和定位作用一样，GIS的空间决策支持作用已逐步应用于ITS的各个服务领域。本文通过对普通GIS空间决策支持作用的分析，提出面向ITS的GIS不仅可以对交通信息起空间定位作用，而且可以运用其对空间数据的分析能力，通过与交通专业的结合，在ITS的各个服务领域起决策支持作用。同时，根据ITS对GIS决策支持的要求，提出了面向ITS的专题GIS应具有的特点。     

东海原甲藻的分子鉴定

采用nrDNAITS序列及18S序列两种分子标记,对东海原甲藻和美国国家海洋藻种保藏中心(CCMP)的具齿原甲藻进行分子鉴定,结果表明,两者之间序列非常相似,两者的nrDNAITS序列碱基差异值仅为0.002,nrDNA18S序列的碱基差异值为0.000,根据分子数据,东海原甲藻和美国国家海洋藻种保藏中心的具齿原甲藻应为同一个种.从基因库获取原甲藻的另外3个种nrD-NAITS序列和8个种的18S序列,用计算机软件进行分析和构建分子系统树,结果显示18S序列用于原甲藻的分子鉴定过于保守,而nrDNAITS更适合于该种属界定的分子标准.     

三株赤潮硅藻5.8S rDNA及转录间隔区(ITS)的克隆及序列分析

对引发赤潮的3株硅藻——1株尖刺拟菱形藻(Pseudo-nitzshia pungens)和2株中肋骨条藻(Skeletonema costatum)的5.8SrDNA和ITS(internal transcribed spacers)序列进行了PCR扩增、克隆和序列测定,并分析了硅藻门10株赤潮藻(7株从GenBank获得)的系统进化关系.研究结果表明,尖刺拟菱形藻的ITS和5.8SrDNA的长度为693bp,SK-1(分离自东海赤潮暴发区)测序得到715bp,除ITS和5.8SrDNA外,还包含部分18SrDNA和28SrDNA;SK-2(分离自青岛养殖场)的ITS和5.8SrDNA的长度为331bp,尖刺拟菱形藻与从GenBank中获得的2株尖刺拟菱形藻相似程度最高,为100%,与该属的多列拟菱形藻相似程度稍低,为82.9%.SK-2的ITS序列与SK-1的相似程度很低,只有51%,但与拟中肋骨条藻的ITS序列相似程度高,为95.5%.SK-1的ITS序列与拟中肋骨条藻的相似程度也低,为56.7%.系统进化树反映的结果与相似性反映的结果一致.研究的该株尖刺拟菱形藻从根据ITS序列研究的结果与形态鉴定的结果看是一致的;SK-2可能属于拟中肋骨条藻;SK-1比较特殊,有待于用其他的方法进一步研究确定其分类地位.     

深圳杨梅坑海域造礁石珊瑚共生藻分类和遗传多样性研究

通过核糖体ITS(Internal Transcribed Spacer)的核苷酸序列研究了深圳杨梅坑海域20种47个造礁石珊瑚样本的共生藻。通过ITS序列分析,与GenBank上的4种不同的共生藻构建Neighbor-Joining聚类树,进行石珊瑚共生藻分类和遗传多样性分析。结果表明,该海域的造礁石珊瑚共生藻属于2种不同的种类(亚系群),19个样品属于C1亚系群共生藻,1个样品属于C15亚系群共生藻,C1和C15两个亚系群共生藻之间的遗传距离为0. 01。将深圳杨梅坑海域得到的ITS序列与NCBI数据库上的福建东山海域、广东徐闻地区的C1系共生藻进行比对,只有深圳杨梅坑海域藻类样本平均(A+T)碱基含量为49. 4%,(A+T)含量小于(G+C)含量。构建Neighbor-Joining聚类树表明,深圳杨梅坑海域石珊瑚共生藻与福建东山海域的亲缘关系较近,而与广东徐闻地区的亲缘关系较远,地理隔离是主要的因素。