东方小藤壶Chthamalus challengeri（甲壳动物亚门：无柄目）线粒体基因组：小藤壶科基因排列比较及藤壶亚目系统发育分析

藤壶是潮间带生态、幼体发育以及生物防污损研究中重要的模式生物。目前,线粒体基因组学的发展有助于从线粒体基因组水平上更好地理解系统发育关系。本研究获得东方小藤壶Chthamalus challengeri完整线粒体基因组,大小为15358bp的环状分子。与藤壶亚目其它物种相比,东方小藤壶非编码区的长度较长,而基因区的长度则相近。东方小藤壶线粒体基因组A+T含量为70.5%。现有藤壶物种的线粒体基因组中存在起始和终止密码子的变化。同属的东方小藤壶和触肢小藤壶C. antennus具有共同的基因排列。然而,小藤壶科中不同属之间却具有不同的基因排列,包括两个tRNA基因出现易位,一个基因块出现倒置。值得注意的是,与以往藤壶亚目物种不同,小藤壶属两个物种的srRNA和lrRNA基因都在重链上编码。小藤壶科中进化树的拓扑结构与基因排列证据相互支持,系统发育分析表明小藤壶科是单系群,而藤壶科和古藤壶科则是多系群。     

藤壶科DNA 分类研究

围胸总目藤壶科的分类系统经历了二亚科系统(小藤壶亚科 Chthamalinae,藤壶亚科 Balaninae)、三亚科系统[藤壶亚科(Balaninae),巨藤壶亚科(Megabalaninae),凹藤壶亚科(Concavinae)],现在采用的是四亚科系统[藤壶亚科(Balaniae)、纹藤壶亚科(Amphibalanus)、巨藤壶亚科(Megabalaninae)和凹藤壶亚科(Concavinae)],但各亚科之间的系统演化关系尚未进行过分子系统学方面的研究.许多藤壶科物种存在趋同进化的趋势,致使传统的形态分类存在困难,不能正确地进行鉴别.本文测定了藤壶科3个亚科里个代表种的线粒体 COI,16S 和12S 基因的部分序列,结合 GenBank 中藤壶科其他物种的12S,28S和18S等基因序列,比较了不同基因片段作为鉴别藤壶科物种的条形码的可行性和有效性,并联合16S和12S序列初步分析了藤壶科各亚科之间的一些亲缘关系.研究结果表明:COI基因的种间和种内遗传距离有明显的间隔区, COI最小种间距离为0.122,远大于最大种内距离0.023,而16S基因的种间与种内距离存在覆盖,最小种间距离为0.018,小于最大种内距离0.023,因此表明,线粒体基因 COI能更准确地鉴定藤壶科种间以及种内关系,并得出阈值为种内差异小于0.023,种间差异大于0.1.ML和BI系统发育分析结果基本一致,支持4亚科的分类系统;巨藤壶亚科形成明显单系群,支持率很高,而两种纹藤壶和管藤壶聚成一支,形成一个单系,本结果支持Newman & Ross的假说,即纹藤壶属和管藤壶属应合并.     

中国近海的蔓足类 Ⅲ.小藤壶科

小藤壶类除少数种为深海产外，大部栖息于潮间带，附着于岩石、码头、树木、管道、浮标或船底，是沿岸附着生物的主要种类，在潮间带岩岸生物群落中常占一定的优势。种类的准确鉴定对分析潮间带生态和附着生物的生态研究及防除工作都具有一定的意义。 早在1854年，Darwin就在藤壶科Balanidae下建立了小藤壶亚科Chthamalinae. Pilsbry (1916)将其提升为小藤壶科Chthamalidae，与Balanidae并列；1976年Newman& Ross对藤壶亚目Suborder Balanomorpha进行了全面的修订，分为3个总科(Chthamaloidea,Coronuloidea和Balanoide)，他们将小藤壶科中壳壁带轮生副板的4属分出，建立鳞藤壶科Catophragmidae，与Chthamalidae一起置于小藤壶总科Chthamaloidea之下。 关于小藤壶类，国内仅有零星记载，缺少系统研究，特别是一些深水种尚无报道。本文根据中国科学院海洋研究所过去在全国沿海潮间带采集的标本写成，共报告小藤壶科9种，分隶于2亚科4属，其中有1新种，1种为我国首次录。新种的模式标本保存在中国科学院海洋研究所。 本文报告的小藤壶，分布于我国长江口以北海域的只有东方小藤壶Chthamalus challengeri Hoek，它大量出现在高潮带岩石上，对温、湿度变化的适应力很强，能忍受较长时间的周期性干燥和较大的温度变化幅度。其余种类则出现在长江口以南亚热带和热带海区，其中白条地藤壶Euraphia withersi和楯形矮藤壶Chamaesipho scutelliformis分布广，数量大；中国四板小藤壶Tetrachthamalus sinensis Ren在海南岛和北部湾沿岸占优势。     

10种蟹类18S rDNA 和COⅠ基因序列分析及其分子系统发育研究

应用分子系统发育学的方法,以蟹类18S rDNA和线粒体细胞色素C氧化酶亚单位Ⅰ(COⅠ)基因序列片段为分子标记,结合形态学特征对10种蟹类的分类地位进行探讨。实验共获得10 条18S rDNA序列,长度为1780~1787 bp,其中A、T, G,C平均含量分别为23.72%, 24.58%,24.52%和27.17%;通过序列对比,发现18S rDNA序列相对保守,只含有1个从88 bp 到130 bp 约 50 bp的高可变区;物种间碱基距离比较小,从0.001到0.017。18S rDNA系统发育树为方蟹总科、沙蟹总科和梭子蟹总科起源于同-海洋蟹类祖先提供了分于生物q证惦,开上火亚N内尔量分别为26.88%.弓蟹科。获得的5条CO Ⅰ基因序列,长度均为709 bp,A、T,G、C平均含量分别为26.88%,37.62%,17.50%和18.00%;COⅠ基因片段变异性高,物种间碱基距离从0.016到0.138。COⅠ基因系统发育树真实地反映了住属各物种之间的进化关系,和传统分类非常吻合,为形态特征相似的姆类鉴定提供了-种快速准确的方法。     

18S rDNA and CO Ⅰ gene sequence analysis of 10 crabs and study of their molecular phylogeny

应用分子系统发育学的方法,以蟹类18S rDNA和线粒体细胞色素C氧化酶亚单位Ⅰ(COⅠ)基因序列片段为分子标记,结合形态学特征对10种蟹类的分类地位进行探讨.实验共获得10条18S rDNA序列,长度为1 780～1 787 bp,其中A、T、G、C平均含量分别为23.72％,24.58％,24.52％和27.17％;通过序列对比,发现18S rDNA序列相对保守,只含有1个从88 bp到130 bp约50 bp的高可变区;物种间碱基距离比较小,从0.001到0.017.18S rDNA系统发育树为方蟹总科、沙蟹总科和梭子蟹总科起源于同一海洋蟹类祖先提供了分子生物学证据,并且支持将厚蟹属从相手蟹科移至弓蟹科.获得的5条CO Ⅰ基因序列,长度均为709 bp,A、T、G、C平均含量分别为26.88％,37.62％,17.50％和18.00％;COⅠ基因片段变异性高,物种间碱基距离从0.016到0.138.CO Ⅰ基因系统发育树真实地反映了归蟳属各物种之间的进化关系,和传统分类非常吻合,为形态特征相似的蟳类鉴定提供了一种快速准确的方法.     

科学有趣的鱼类分类

地球上的鱼类大约有2万多种。如何将它们分门别类地区别开来,这既是一个包含生物分类科学的严谨工作,又是一个引人人胜的话题。我们知道,现代分类学上(包括对鱼的分类)采用的等级主要有门、纲、目、科、属、种,必要时还可以补充一些等级,如亚门、总纲、亚纲、总目、亚目、总科、亚科、亚属等。某种生物作为物种是真实存在的,并不是人为地分类划分。自然界有形形色色的各种生物,在大多数情况下,物种之间有明确的界     

中国常见贻贝基于28S rDNA序列的系统发育分析

贻贝隶属于软体动物门(Mollusca)、双壳纲(Bivalvia)、翼形亚纲(Pteriomorphia)、贻贝目(Mytilida)、贻贝超科(Mytiloidea),大约有400种贻贝分布在世界各地,可适应淡水、潮间带至深海多种生境。本实验以贻贝科6亚科12属28种中国沿海常见贻贝的28SrDNA为目的片段,构建系统发育进化树,运用最大似然法和贝叶斯推演法分析了贻贝科的系统发生,并追踪贻贝科物种系统演化历史。结果显示:贻贝亚科Mytilinae、偏顶蛤亚科Modiolinae、石蛏亚科Lithophaginae均非单系群。在属阶元,深海偏顶蛤属Bathymodiolus、贻贝属Mytilus和股贻贝属Perna为单系群。本研究发现应接受将原隔贻贝属Septifer分为Septifer属和Mytilisepta属的分类提议;应接受将原石蛏属Lithophaga中的膜石蛏亚属Leiosolenus提升至属的地位的分类提议。此外,短齿蛤属Brachidontes的单系性不被支持,刻缘短齿蛤Brachidontessetiger并未与短齿蛤属其他物种在系统发育树上聚拢,亲缘关系较远,为不同属物种,建议恢复刻缘短齿蛤原属名Volsella (Dunker, 1857)。     

寄生亚目的系统研究概况以及我国寄生亚目的研究展望

寄生亚目（Epcaridea）隶属于节肢动物门（Arthropoda）甲壳动物亚门（Crustacea）软甲纲（Malacostraca）等足目（Isopoda）,成体是甲壳类的专性外寄生动物,多附生于十足目的鳃室或腹肢间的腹甲表面。本亚目多为海生种类,极少数为淡水生活。大多数种在找到宿主前,先经历浮游期。据Kensley和Schotte统计寄生亚目有近800种,隶属于2总科11科约304属。关于寄生亚目的分类系统,学者们的争议较大。Bonnier主要根据寄主的不同,将寄生亚目分为2部分Bopyrinae和Cryptoniscinae,前者包含4科,分别为檐虱科Dajidae Giard＆ Bonnier,1887,软虱科Phryxidae Bonnier,1900,鳃虱科Bopyridae Rafinesque,1815,内虱科Entoniscidae Kossmann,1881;后者包含8科,分别为微虱科Microniscidae Bonnier,1900,半虱科Hemioniscidae Bonnier,1900,Cyproniscidae Bonnier,1900,Liriopsidae Bonnier,1900,囊虱科Asconiscidae Bonnier,1900,毛虱科Crinoniscidae Bonnier,1900,足虱科Podasconidae Bonnier,1900,Cabiropsidae Giard＆ Bonnier,1887。     

基于16S rRNA 部分序列探讨部分鳚亚目鱼类的分子系统进化关系

通过PCR扩增获得了3科5属6种中国黄渤海海域的鳚亚目(Blennioidei)鱼类的线粒体16S rRNA基因序列片段约669bp碱基,结合来自 GenBank的5种鳚亚目其他科鱼类的相应基因片段,并以眼斑雪冰鱼(Chionodraco rastrospinosus)为外群,生成供系统发育分析的序列矩阵,利用MEGA 4.0软件分析序列的碱基组成、差异百分比、转换/颠换值等,应用最大简约法(MP)和邻接法(NJ)构建系统发育树.结果显示:在鳚亚目鱼类的16S rRNA 片段生成的序列矩阵中发现有碱基的插入缺失现象,共有207 bp变异位点,转换/颠换值为0.8,碱基平均差异为3.36;支持绵鳚(Enchelyopus elongates)归于鳚亚目绵鳚科(Zoarcidae),鳚(Azuma emmnion)归于鳚亚目线鳚科(Stichaeidae);方氏云鳚(Enedrias fangi)和云鳚(Enedrias nebulosus)种间遗传距离只有0.01,亲缘关系最近.     

多毛类18S rDNA和GOⅠ基因序列片段及其分子系统发育研究

应用分子系统发育学的方法,以多毛类18S rDNA和线粒体细胞色素C氧化酶亚单位Ⅰ(CO Ⅰ)基因序列片段为分子标记,结合形态学特征对它们的分类地位进行了探讨.结果显示,四索沙蚕-梯斑海毛虫分枝的遗传距离为0.044,(四索沙蚕-梯斑海毛虫)-小头虫分枝的遗传距离为0.072,四索沙蚕分枝与岩虫分枝的遗传距离比较大,为0.091.分析结果显示18S系统发育树将仙女虫科定位为矶沙蚕目的内群分类单元,100%的Bootstrap值进一步表明仙女虫科和索沙蚕科的姐妹群关系;分析结果还显示了小头虫目与矶沙蚕目的亲缘关系密切.     

Microbial Biomass and Organic Nutrients in the Deep-Sea Sediments of the Central Indian Ocean Basin

In order to assess the impact of deep-sea mining on the in situ benthic life, we measured the microbial standing stock and concentration of organic nutrients in the deep-sea sediments of the Central Indian Ocean Basin in the Indian pioneer area. Sediments were collected using box core and grab samples during September 1996. The total bacterial numbers ranged from 10 10 -10 11 cells per g -1 dry weight sediment. There was a marginal decrease in the number of bacteria from surface to 30 cm depth, though the subsurface section registered a higher number than did the surface. The highest numbers were encountered at depths of 4-8 cm. The retrievable number of bacteria were two orders less in comparison with the direct total counts of bacteria. An almost homogeneous distribution of bacteria, total organic carbon, living biomass, and lipids throughout the depth of cores indicates active microbial and benthic processes in the deep sea sediments. On the other hand, a uniform distribution of total counts of bacteria, carbohydrates, and total organic carbon in all the cores indicates their stable nature and suggests that they can serve as useful parameters for long-term monitoring of the area after the benthic disturbance. Further studies on temporal variability in this region would not only verify the observed norms of distribution of these variables but would also help to understand restabilization processes after the simulated benthic disturbance.     

Assessing the Validity of Seismo-Acoustic Predictor Equations for Obtaining Seabed and Sub-Surface Sediment Physical Properties

The interdependence between the seismo-acoustic properties of a marine sediment and its geotechnical/physical parameters has been known for many years, and it has been postulated that this should allow the extraction of geotechnical information from seismic data. Though in the literature many correlations have been published for the surficial layer, there is a lack of information for greater sediment depths. In this article, a desktop study on a synthetic seafloor model illustrates how the application of published near-surface prediction equations to subsurface sediments (up to several tens of meters burial depth) can lead to spurious predictions. To test this further, acoustic and geotechnical properties were measured on a number of sediment core samples, some of which were subjected to loading in acoustically-equipped consolidation cells (oedometers) to simulate greater burial depth conditions. For low effective pressures (representing small burial depths extending to around 10 meters subsurface), the general applicability of established relationships was confirmed: the prediction of porosity, bulk density, and mean grain size from acoustic velocity and impedance appears generally possible for the investigated sedimentary environments. As effective pressure increases through, the observed relationships deviate more and more from the established ones for the near-surface area. For the samples tested in this study, in some instances increasing pressure even resulted in decreasing velocities. There are several possible explanations for this abnormal behavior, including the presence of gas, overconsolidation, or bimodal grain size distribution. The results indicate that an appropriate depth correction must be introduced into the published prediction equations in order to obtain reliable estimates of physical sediment properties for greater subsurface depths.     

A Review of Some Environmental Issues Affecting Marine Mining

This article reviews information recently available from existing marine and coastal mining for responses to environmental issues affecting marine mining at different depths. It is particularly but not exclusively concerned with those issues affecting seabed biodiversity impact and recovery. Much information has been gathered in the past 10 years from shallow mining operations for construction aggregate, diamonds, and gold, from coastal mines discharging tailings to shallow and deep water, and from experimental deep mining tests. The responses to issues identified are summarized in a series of eight tables intended to facilitate site-specific consideration. Since impacts can spread widely in the surface mixing layer SML, and can affect the biologically productive euphotic zone, the main issues considered arise from the depth of mining relative to the SML of the sea. Where mining is below the SML, the issue is whether it is environmentally better to bring the extraction products to the surface vessel for processing (and waste discharge), or to process the extraction products as much as possible on the seabed. Responses to the issues need to be sitespecific, and dependent on adequate preoperational environmental impact and recovery prediction. For deep tailings disposal from a surface vessel, there are four important environmental unknowns: (1) the possible growth of "marine snow" (bacterial flocs) utilizing the enormous quantities of fine tailings particles (hundreds or thousands of metric tons per day) as nuclei for growth, (2) the possibility that local keystone plankton and nekton species may migrate diurnally down to and beyond the depth of deep discharge and hence be subjected to tailings impact at depth, (3) the burrow-up capability of deep benthos and their ability to survive high rates of tailings deposition, and (4) the pattern and rate of dispersion of a tailings density current through the deep water column from discharge point to seabed. Actions to obtain relevant information in general and site-specifically are suggested.     

Increased Particle Fluxes at the INDEX Site Attributable to Simulated Benthic Disturbance

Particle fluxes were measured 7 m above the sea bottom during the predisturbance, disturbance, and postdisturbance periods by using time series sediment traps attached to seven deep-sea moorings deployed in the INDEX experiment site in the Central Indian Basin. The predisturbance particle fluxes varied between 22.3 to 55.1 mg m -2 day -1 . Increased and variable particle fluxes were recorded by the sediment traps during the disturbance period. The increase observed was 0.5 to 4 times more than the background predisturbance fluxes. The increases in particle fluxes (~4 times) recorded by the sediment trap located in the southwestern direction (DMS-1) were the greatest, which could be the result of preferential movement of resuspended particles generated during the deep-sea benthic disturbance along the general current direction prevailing in this area during the experimental period. Also, the traps located closer to the disturbance area recorded greater fluxes than did the traps far away, across the Deep Sea Sediment Resuspension System path. This variability in recorded particle fluxes by the traps around the disturbance area clearly indicates that physical characteristics such as grain size and density of the resuspended particles produced during the disturbance had an important effect on particle movement. The postdisturbance measurements during ~5 days showed a reduction in particle fluxes of ~50%, indicating rapid particle settlement.     

Geoacoustic Inversion via Genetic Algorithm and Its Application to Manganese Sediment Identification

An acoustic inversion method using a wide-band signal and two near field receivers is proposed and applied to multiple layered seabed models including a manganese sediment. The inversion problem can be formulated into a probabilistic model comprised of signals, a forward model, and additive noise. The forward model simulates wide-band signals, such as chirp signals, and is chosen to be the source-waveletconvolution plane wave modeling method. The wavelet matching technique, using weighted least-squares fitting, estimates the sediment sound-speed and thickness on which determination of the possible numerical ranges for a priori uniform distribution is based. The genetic algorithm is applied to a global optimization problem to find a maximum a posteriori solution for determined a priori search space. Here the object function is defined by an L 2 norm of the difference between measured and modeled signals. Not only the marginal pdf but also its statistics are calculated by numerical evaluation of integrals using the samples selected during importance sampling process of the genetic algorithm.     

海上大直径钢管桩打桩过程中桩周土强度弱化研究

海上大直径钢管桩打桩过程中,桩周土体受到强烈扰动而发生强度弱化,掌握桩周土体强度弱化规律对于准确预测打桩过程、保证工程安全具有重要意义。为研究土体强度弱化规律,开展了环剪试验模拟打桩对桩周土体的扰动,测试土体强度随剪切速率的变化规律,建立了描述土体强度弱化规律的拟合公式,引入到打桩分析软件中。研究结果表明：土体的强度折减程度不仅与土体本身的性质有关还受到土体的埋深和剪切速率的影响,埋深越深土体强度折减程度越低,剪切速率越高土体强度折减越高,在打桩分析中可采用这里推荐的线性折减方法来模拟不同深度处土体强度的折减规律。     

Determination of In Situ Sediment Shear Strength from Advanced Piston Corer Pullout Forces

The advanced piston corer (APC) has been used by the Ocean Drilling Program since 1985 for recovering soft sediments from the ocean floor. The pullout force measured on extracting the core barrel from the sediment is shown to correlate with the average shear strength of the sediment core measured in the ship's laboratory. A simple rule of thumb is derived relating the shear strength of the sediment to the pullout force. Multiple APC holes at individual sites allow the consistency of the pullout measurements to be assessed. The effects of different operational procedures during APC coring are also explored. Although generally applicable, the correlation between pullout force and laboratory measurements of shear strength breaks down for some APC holes, possibly because of the disturbance of some sediment types during the APC coring process. A better understanding of the physical process of APC coring, and its effect on the properties of the sediment both inside and immediately outside the core barrel, would indicate what confidence can be put on the measurement of pullout force as a way of evaluating the in situ shear strength of deep sea sediments.     

Interannual Sea Level Variations and Annual Tides in the Northwestern Pacific

Specific properties of the interannual sea level variations and annual tides in the Northwestern Pacific were studied. Several tide stations were monitored. The monthly mean sea level for the year of 1995 was analyzed at each tide station. A seismic event in 1995, some tectonic activity around the subject area, and the Kuroshio (the oceanic western boundary current) may possibly contaminate results which would have occurred from the astronomical annual tide alone.     

Flow Cytometric Analysis of Coastal Lagoon Bacterioplankton and Picophytoplankton: Fixation and Storage Effects

The effect of different fixation and storage protocols on the flow cytometric (FCM) simultaneous analysis of bacterioplankton and phytoplankton in coastal seawater samples (Mediterranean coastal lagoons) was investigated. FCM measurements (cell number, fluorescence and scatter characteristics) were obtained through DAPI staining. Three fixatives [glutaraldehyde (GA), formaldehyde (FA) and paraformaldehyde (PFA)] and two storage (3 months duration) methods (5 °C and −196 °C) were tested. Two dominant populations were detected in studied samples: bacteria and eukaryotic picophytoplankton. Adding fixatives (2% final concentration) appears necessary to obtain FCM exhaustive counts of all the bacteria and phytoplanktonic cells. This was related to the permeation effect of fixatives which allowed a better DAPI staining of the cells. Maximum fluorescence, i.e. optimal staining of the cells was obtained with FA or PFA, and significant lower fluorescences with GA. Fixed samples stored at 5 °C induced rapid cell loss. Only storage in liquid nitrogen of samples fixed with FA or PFA, allows mid-term (≥4 months) preservation of bacteria or picophytoplankton cell numbers, and limited evolution of DAPI-induced fluorescence and scatter characteristics.