大型溞(Daphnia magna)线粒体基因组的测定与序列分析

利用Long-PCR及普通PCR技术,结合引物步移法测序,经拼接、组装后获得中国淮河大型溞(Daphnia magna)线粒体基因组全序列,并对其基因组成、结构特点等进行初步分析.大型溞线粒体基因组全长为14948 bp,A、T、G、C各碱基含量分别为32.37%、34.73%、15.58%、17.31%,表现出明显的AT偏倚.13个蛋白质编码基因、22个t RNA基因、2个rRNA基因和1个D-loop区(控制区)的排列方式与典型的节肢动物线粒体基因组的排列方式略有不同.除COI基因以CTG、ATP8基因以GTG、ND3基因以ATC以及ND6基因以ATT为起始密码子以外,其余9个蛋白质编码基因均以ATG作为起始密码子;9个蛋白质编码基因具有典型的完全终止子TAG或TAA,COI、COII、ND4和ND5等基因采用不完全终止密码子T.基因组包含9个基因间隔区,共81 bp,长度1~62 bp;13个基因重叠区,共77 bp,重叠碱基数在1~27 bp之间,最大重叠在16 S rRNA和tRNA~(Val)基因之间.在预测的22个tRNA基因的二级结构中,发现只有tRNA~(Ser1)基因未能形成完整的二级结构,其他21个基因均可形成典型的三叶草结构.16 S rRNA和12 S rRNA基因长度分别为1373 bp和752 bp,D-loop区全长289 bp.本研究结果为探讨大型溞在溞属中的系统学地位及其与其他种间的系统发生关系等问题提供了数据资源.     

5个湖泊河川沙塘鳢(Odontobutis potamophila)种群线粒体细胞色素b基因的遗传变异分析

通过对5个湖泊的河川沙塘鳢种群的线粒体DNA细胞色素b基因进行PCR扩增、测序,获得1141 bp的序列全长.序列分析显示,cyt b基因序列中A+T含量(55.8%)略高于G+C含量(44.2%),共检测到806个多态位点,115个样本得到87个单倍型,平均单倍型多样性为0.969±0.012,核苷酸多样性为0.20081±0.00742,遗传多样性表现高度多样性.太湖种群与大纵湖种群间的遗传距离最近,为0.137,巢湖种群和大纵湖种群之间遗传距离最远,为0.424.分子方差分析表明,群体间遗传分化系数Fst为0.531,变异来自群体内及群体间.cyt b基因序列构建的UPGMA系统进化树显示,5个种群分化成不同的分支系谱,种群间存在的基因交流较少.     

湖泊底泥中微囊藻DNA的分子检测

通过改进裂解温度和延长裂解时间并增加苯酚/氯仿洗脱次数的DNA提取方法获得南京玄武湖底泥中的DNA,通过PCR法来扩增微囊藻的16SrRNA基因.结果表明在所有采样点中均得到微囊藻基因组DNA,并且纯度较高,OD260/OD280均高于1.54,最高值达到1.89.PCR的扩增结果显示所有样点的DNA都得到212 bp大小的微囊藻16SrRNA基因片断,表明这种方法可以有效的从底泥中提取微囊藻的DNA,从而为研究底泥微囊藻生理生态及其越冬、上浮、形成水华的机理提供更有利的方法.     

五个湖泊中华刺鳅(Mastacembelus sinensis)种群线粒体细胞色素b遗传多样性分析

采用特异性引物对中华刺鳅基因组DNA进行扩增和测序,获得了江苏省5个湖泊的中华刺鳅种群线粒体cyt b基因767 bp的序列.分析表明,cyt b基因中A+T含量略高于G+C含量,144个中华刺鳅共获得cyt b基因单倍型28个,变异位点38个,其中单倍型Hap1分布最为广泛,可能为祖先型单倍型.平均单倍型多样性和平均核苷酸多样性指数分别为0.58751和0.00214,遗传多样性较低.群体间遗传分化系数为0.0356,绝大多数的遗传变异都来自群体内部,而群体间的遗传分化极小.构建的系统进化树未出现明显的以单个湖泊群体的聚簇,5个湖泊的种群间基因交流极其强烈.研究结果表明,当前5个湖泊的中华刺鳅遗传多样性水平比较低,亟待加强保护.     

长江中下游5个湖泊黄颡鱼(Pelteobagrus fulvidraco)种群线粒体细胞色素b基因的遗传变异分析

对长江中下游5个湖泊的黄颡鱼种群的线粒体DNA细胞色素b基因进行了PCR扩增、测序,获得了955 bp的序列.分析显示,cyt b序列中A+T含量略高于G+C含量,共检测到54个变异位点,60个样本得到37个单倍型,平均单倍型多样性为0.945±0.018,核苷酸多样性为0.00419±0.00043,遗传多样性表现中等.太湖种群与滆湖种群间的遗传距离最远为0.00651,鄱阳湖种群和巢湖种群之间遗传距离最近为0.00375.分子方差分析(AMOVA)表明,群体间遗传分化系数F_st为0.0684,几乎所有变异都来自群体内,群体间遗传分化极小.cyt b序列构建UPGMA系统进化树显示,5个种群没有分化成不同的分枝谱系,种群间存在广泛的基因交流.     

微囊藻胶鞘多糖和水溶性胞外多糖的化学特性

微囊藻(Microcystis)产生大量胞外多糖(EPS),包括包裹在细胞外的胶鞘多糖(CPS)和释放到周围环境中的水溶性多糖(RPS).为探究EPS在蓝藻水华发生中的生理生态学意义,迫切需要了解微囊藻EPS的化学特性.本文从太湖分离群体微囊藻,经过大约18个月实验室培养后,其中一些藻株转变为单细胞形态.选择5株群体藻和4株单细胞藻,比较分析这些藻株EPS的化学特性发现:(1)所有9株藻的EPS均为含有脱氧己糖的酸性杂多糖;(2)所有9株藻的CPS的糖醛酸含量(1.2%~2.1%)均低于RPS的糖醛酸含量(2.4%~6.2%);(3)所有9株藻的EPS均含有乙酰基和硫酸基,其中,每一株藻CPS的乙酰基含量均高于RPS的乙酰基含量,所有群体藻CPS的乙酰基含量(4.1%~6.6%)高于所有单细胞藻CPS的乙酰基含量(2.0%~3.2%).本文进而讨论了EPS化学特性对EPS水溶性和微囊藻群体形成的影响,以及对其生态学作用的影响.在这些化学特性中,乙酰取代基团被认为可能是影响微囊藻EPS生理生态学作用的重要因素.     

汾河(太原市景区段)微囊藻的分子多样性及产毒能力

对太原市汾河景区采集到的微囊藻进行分离纯化,得到7株微囊藻藻株,运用ITS、PC-IGS和gyrB基因序列构建系统树来研究其分子多样性.结果表明,太原市汾河景区微囊藻具有一定的分子多样性.采用mcyA基因和ELISA检测两种方法,即全细胞PCR测定微囊藻毒素合成酶基因A( mcyA),对这些藻株的微囊藻毒素进行测定,检...     

角突网纹溞在太湖微囊藻群体形成中的作用

将太湖微囊藻水华中3种优势微囊藻包括水华微囊藻1028、惠氏微囊藻929、铜绿微囊藻469和铜绿微囊藻905培养在改良后的BG-11培养基(TN=10mg/L,TP=0.4mg/L)中,然后加入角突网纹搔,以研究3种优势微囊藻对浮游动物摄食压力的形态反应.整个实验共进行了12d.除了水华微囊藻1028以外,在惠氏微囊藻929、铜绿微囊藻469和铜绿微囊藻905中没有观察到有大群体(大于10个细胞)的出现.在水华微囊藻中,处理组大群体细胞所占总细胞的比例与对照组显著不同,其中对照组占22%,而试验组占53%.水华微囊藻对照组和处理组中单细胞、2细胞、小群体(3-10个细胞)和大群体(大于10个细胞)细胞密度存在显著的不同.实验第6-12d,水华微囊藻对照组和试验组单位大群体细胞数量存在显著差异.研究结果表明,角突网纹潘的摄食压力不能促使惠氏微囊藻929、铜绿微囊藻469和铜绿微囊藻905形成大群体.角突网纹溞的摄食促使水华微囊藻形成更大的群体.     

四个鲤鱼种群ITS-1序列的遗传变异分析

对我国四个鲤鱼种群的核糖体DNA内转录间隔区ITS-1进行了PCR扩增、测序.序列分析显示,370bp的ITS-1序列中GC含量明显高于AT含量,共检测到34个变异位点,96个样本得到14个单倍型,平均单倍型多样性为0.637±0.055,核苷酸多样性为0.00857±0.00200,遗传多样性表现较低.黑龙江野鲤种群...     

鳙鱼微囊藻毒素去毒酶基因cDNA全序列的克隆与序列分析

淡水鱼类可溶性谷胱甘肽S-转移酶(soluble glutathione S-transferase,sGST)在微囊藻毒素去毒代谢过程中起着关键作用,又被称为微囊藻毒素去毒酶.通过简并引物克隆鳙鱼肝脏sGST基因cDNA核心序列,应用5'-RACE和3'-RACE技术分别扩增该序列的5'末端和3'末端序列,最后通过序列拼接获得鳙鱼肝脏sGST基因cDNA全序列.序列分析结果表明,鳙鱼肝脏sGST基因cDNA全长934 bp,其中5'-UTR长104bp,3'-UTR长158 bp,编码区672 bp,编码223个氨基酸.鳙鱼与海水鱼类、鸟类、哺乳类等不同类群sGST的N-末端功能域氨基酸同源性较高,达75%左右,而C-末端功能域氨基酸同源性较低,仅为43.6%-55.9%,这与脊椎动物sGST的N-末端功能域均存在相同的GSH结合位点,而C-末端功能域存在不同的底物结合位点相一致.鳙鱼sGST的C-末端功能域的低同源性可能与其承担微囊藻毒素去毒代谢之特殊功能有关.     

Detection of Toxoplasma gondii in Turkish River and Drinking Water Samples by Different PCR and LAMP Methods

In this study, we investigated the occurence of Toxoplasma gondii and checked the sequencing in 56 samples collected from in the Ordu province, Middle Black Sea, Turkey. The samples were collected from drinking and river water and DNA was extracted from all samples. After DNA isolation, the 18S rRNA target gene and B1 gene of T. gondii were amplified by conventional PCR, nested PCR, and loop‐mediated isothermal amplification (LAMP) assay, respectively. Twenty out of 56 samples (35.7%) were Toxoplasma positive by LAMP. The conventional PCR and nested PCR confirmed that 12 and 16 out of 56 samples (21.42 and 28.57%) were positive for Toxoplasma, respectively. All drinking water samples were negative by all three methods. Thirteen Toxoplasma nested PCR products were successfully sequenced. This study achieved detection and sequencing of Toxoplasma from water supplies in the Middle Black sea area in Turkey.     

A Sensitive and Versatile Multiplex PCR System for the Rapid Detection of Enterotoxigenic (ETEC), Enterohaemorrhagic (EHEC) and Enteropathogenic (EPEC) Strains of Escherichia coli

Although Escherichia coli is widely distributed in the marine environment, only a small percentage are pathogenic to humans. Nonetheless, the widespread occurrence of waterborne infections of E. coli origin in humans has become one of the major health problems worldwide. To date, several types of enterovirulent E. coli have been recognized as the aetiologic agents of various gastrointestinal infections in humans. The most commonly encountered are those belonging to the enterotoxigenic (ETEC), enteroinvasive (EIEC), enterohaemorrhagic (EHEC) and enteropathogenic (EPEC) subtypes. In order to better determine the health risks that are associated with exposure to some of these specific subtypes, we have developed a very sensitive multiplex PCR system for the rapid detection and typing of ETEC, EHEC and possibly EPEC strains of E. coli in the aquatic environment. The target genes chosen for this investigation included: the PHO-A housekeeping gene (present in all E. coli); the LT1, LT2 and ST1 genes of ETEC; the VT1 and VT2 verotoxin, and EAE virulence genes of EHEC and EPEC, respectively. Six pairs of oligonucleotide primers were designed to simultaneously amplify internal fragments of these genes by multiplex PCR to generate PCR products that could be analysed and confirmed with relative ease by gel electrophoresis and HincII enzyme digestion. The results showed that the six sets of PCR primers were highly specific for their target genes and produced specific amplimers of the expected size from several control strains of E. coli – ATCC 35401 (LT1+/ST1+); SA53 (LT2+/VT2+); and O157 (VT1+/VT2+/EAE+). The detection sensitivity of the multiplex PCR system for the six target genes in an E. coli cell mixture was optimized and enhanced by preincubating serially diluted cells in Luria-Bertani broth for 6 h prior to PCR analysis. The results obtained indicated a detection sensitivity of 10° CFU (of each strain) per 100 μl reaction. Multiplex PCR analysis of seawater samples collected from four sewage-polluted sites in Hong Kong indicated the presence in all four samples of E. coli bacteria that were positive for LT1, ST1, VT1 and EAE virulence genes. Overall, the data indicated that the multiplex PCR system described in this study is a potentially very useful and powerful method for routine monitoring and risk assessment of water quality.     

大空间单跨门式刚架结构抗火性能影响因素的客观权重研究

火灾时的大空间单跨门式刚架结构抗火性能,受钢构件温升和结构整体抗火临界温度的影响。本文通过对无保护大空间建筑钢构件温升数据的统计分析,获得包含火源功率、建筑体积、钢构件截面形状系数的钢构件温升拟合公式;结合单跨门式刚架结构整体抗火临界温度数据的统计分析,获得包含荷载比、柱高和系数K的结构整体抗火临界温度拟合公式;通过分析上述因素对钢构件温度变化率及抗火临界温度变化率的影响,获得各个因素的客观权重值,并应用于具体案例的结构安全分析。结果表明,在本文的研究条件下,荷载比权重值最大,约为0.4;火源功率是仅次于荷载比的一个重要因素;建筑体积、截面形状系数和系数K三者都在其自身值较小时,权重值较大,且都随着其自身值的增加,权重值降低。本研究可供无保护门式刚架结构建筑的抗火设计、火灾风险评估和消防管理作参考。     

Application of a mixing-ratios based formulation to model mixing-driven dissolution experiments

We address the question of how one can combine theoretical and numerical modeling approaches with limited measurements from laboratory flow cell experiments to realistically quantify salient features of complex mixing-driven multicomponent reactive transport problems in porous media. Flow cells are commonly used to examine processes affecting reactive transport through porous media, under controlled conditions. An advantage of flow cells is their suitability for relatively fast and reliable experiments, although measuring spatial distributions of a state variable within the cell is often difficult. In general, fluid is sampled only at the flow cell outlet, and concentration measurements are usually interpreted in terms of integrated reaction rates. In reactive transport problems, however, the spatial distribution of the reaction rates within the cell might be more important than the bulk integrated value. Recent advances in theoretical and numerical modeling of complex reactive transport problems [De Simoni M, Carrera J, Sanchez-Vila X, Guadagnini A. A procedure for the solution of multicomponent reactive transport problems. Water Resour Res 2005;41:W11410. doi: 10.1029/2005WR004056, De Simoni M, Sanchez-Vila X, Carrera J, Saaltink MW. A mixing ratios-based formulation for multicomponent reactive transport. Water Resour Res 2007;43:W07419. doi: 10.1029/2006WR005256] result in a methodology conducive to a simple exact expression for the space–time distribution of reaction rates in the presence of homogeneous or heterogeneous reactions in chemical equilibrium. The key points of the methodology are that a general reactive transport problem, involving a relatively high number of chemical species, can be formulated in terms of a set of decoupled partial differential equations, and the amount of reactants evolving into products depends on the rate at which solutions mix. The main objective of the current study is to show how this methodology can be used in conjunction with laboratory experiments to properly describe the key processes that occur in a complex, geochemically-active system under chemical equilibrium conditions. We model three CaCO₃ dissolution experiments reported in Singurindy et al. [Singurindy O, Berkowitz B, Lowell RP. Carbonate dissolution and precipitation in coastal environments: Laboratory analysis and theoretical consideration. Water Resour Res 2004;40:W04401. doi: 10.1029/2003WR002651, Singurindy O, Berkowitz B, Lowell RP. Correction to Carbonate dissolution and precipitation in coastal environments: laboratory analysis and theoretical consideration. Water Resour Res 2005;41:W11701. doi: 10.1029/2005WR004433], in which saltwater and freshwater were mixed in different proportions. The integrated reaction rate within the cell estimated from the experiments are modeled independently by means of (a) a state-of-the-art reactive transport code, and (b) the uncoupled methodology of [12, 13], both of which use dispersivity as a single, adjustable parameter. The good agreement between the results from both methodologies demonstrates the feasibility of using simple solutions to design and analyze laboratory experiments involving complex geochemical problems.     

Hazard and risk assessment in a complex multi-source volcanic area: the example of the Campania Region, Italy

In order to zone the territory of Campania Region (southern Italy) with regard to the hazard related to future explosive activity of Somma-Vesuvio, Campi Flegrei, and Ischia Island, we drew a multi-source hazard map for tephra and pyroclastic flows. This map, which merges the areas possibly endangered by the three volcanic sources, takes into account a large set of tephra fall and pyroclastic flow events that have occurred in the last 10 ka. In detail, for fall products at Campi Flegrei and Somma-Vesuvio we used the dispersal of past eruption products as deduced by field surveys and their recurrence over the whole area. For pyroclastic flows, the field data were integrated with VEI = 4 simulated events; about 100 simulations sourcing from different points of the area were performed, considering the different probability of vent opening. The spatial recurrence of products of both past eruptions and simulated events was used to assign a weight to the area endangered by the single volcanic sources. The sum of these weights in the areas exposed to the activity of two sources and/or to different kinds of products was used to draw a hazard map, which highlights the spatial trend and the extent of the single equivalent classes at a regional scale. A multi-source risk map was developed for the same areas as the graphic result of the product of volcanic hazard and exposure, assessed in detail from a dasymetric map. The resulting multi-source hazard and risk maps are essential tools for communication among scientists, local authorities, and the public, and may prove highly practical for long-term regional-scale mitigation planning.     

河北省40年防震减灾工作回顾与展望--纪念邢台地震40周年

40年来,河北的防震减灾工作在中国地震局和省委、省政府的领导下,全省地震工作者和各有关部门,为建立、推进和发展我省的防震减灾事业做出了不懈的努力和探索,特别是“九五”、“十五”以来,我省认真贯彻落实党和国家关于防震减灾的各项方针、政策,在加强领导、建立健全防震减灾工作体系,加大投入、夯实监测预报基础,加强综合防御、依法推进防震减灾工作,发挥政府职能作用、切实做好地震应急准备等方面,取得了一些进步与成绩。认真、全面、科学地对河北省40年来的防震减灾工作进行回顾总结,对于我们发扬成绩、开创未来,具有重要的意义。     

A simple method of magnetic declination fluctuation correction for long single-streamer positioning in a seismic survey

In single‐streamer acquisition, the use of acoustic transducers to constrain the receiver positions is not possible, thus implying the use of compass birds to gather information on the streamer location. The compasses are, however, sensitive to the declination of the local magnetic field of the earth, and local fluctuations not accounted for can degrade the accuracy of reconstructed positions. In order to correct these small‐scale fluctuations, we propose a simple deterministic method to calculate a spatial correction to apply to the compasses that enhances the positioning accuracy. The local compass declination is calculated after a first reconstruction on the whole survey area. This method was applied with success to navigation data from a 3D survey offshore Japan, and the positioning accuracy was improved to the level of the DGPS accuracy.     

无线数字遥测地震台网的信道测试

在无线数字遥测地震台网中,信道测试是台网传输的关键环节,关系到台网的运行率和地震信号的可用率,甚至整个台网建设的成败。信道测试有3方面的内容:中继站背景场强测试、子台信道储备测试和信道误码测试。经测试后得到各种参数柱状图,以此判断每个台站的信道质量情况,及时调整台网信道传输的设计方案,确保成功建成无线数字遥测地震台网。     

基于COI基因序列的太湖新银鱼遗传多样性

利用线粒体细胞色素C氧化酶I(COI)分子标记分析长江中下游太湖新银鱼(Neosalanx taihuensis)8个地理种群132个样本的遗传多样性.该基因630 bp片段的碱基序列共检出8个核苷酸变异位点(变异率1.27%),其中局域性单倍型居多(75%),群体单倍型多样性较高(h=0.576±0.036),而核苷酸多样性较低(π=0.00112±0.00204).不同地理种群遗传多样性差异显著:有人工移植历史种群遗传多样性较高、隔离度较高的种群遗传多样性较低,但大部分的遗传变异来自于种群内(54.83%),反映出地理隔离和人为干扰对太湖新银鱼遗传格局影响显著.研究表明COI基因适于银鱼科鱼类物种鉴别和系统发育研究,同时可为同种种群间遗传关系分析提供一定的信息.