紫红笛鲷遗传多样性的AFLP分析

针对广东、海南省4个不同地方(阳江、湛江、榆林、琼海)网箱养殖的紫红笛鲷(Lutjanus argentimaculatus)群体和在三亚采集的野生紫红笛鲷群体,利用AFLP分子标记技术分析我国南方养殖的紫红笛鲷的遗传多态性。最适合的AFLP引物组合是E AGC／M CAA,扩增出的AFLP特征位点为107,养殖样品扩增出的AFLP的带数在5474之间,野生样品扩增出的AFLP的带数为63,4个养殖点紫红笛鲷的多态性标记百分率都超过了40．00％,呈现一定程度的遗传多样性。总的来讲,4个养殖地的紫红笛鲷群体之间的遗传距离差异较小,但与野生群体的遗传距离相对较大,UPGMA聚类分析明显将养殖群体与野生群体分开,表明紫红笛鲷养殖群体已经与野生群体存在一定程度的遗传隔阂。     

半滑舌鳎（Cynoglossus semilaevis）野生与养殖群体遗传多样性的比较研究

采用扩增片段长度多态性技术（AFLP技术）对2个野生和2个养殖的半滑舌鳎群体进行了遗传多样性的比较研究。实验采用8对AFLP引物组合,在四个群体的120个个体中共扩增出498个位点。结果表明,半滑舌鳎养殖群体的遗传多样性降低：河北唐山野生群体和江苏连云港野生群体的多态位点百分数分别为45．18％和39．96％,Nei遗...     

用RAPD方法对不同色三疣梭子蟹的分析

为寻找三疣梭子蟹体色相关的分子标记,本文用RAPD方法对紫色和茶绿色两种三疣梭子蟹基因组DNA进行体色相关遗传标记的筛选及其遗传多样性分析。从80个随机引物中共筛选了30个重复性好多态性高的引物,RAPD分析结果表明：紫色梭子蟹个体组成的群体多态性位点比例为61.29%,Nei氏基因多样性指数为0.3756,Shannon信息指数为0.5545;茶绿色梭子蟹个体组成的群体多态性位点比例为62.44%,Nei氏基因多样性指数为0.3451,Shannon信息指数为0.5099。紫色和茶绿色三疣梭子蟹之间Nei氏无偏差遗传相似性指数为0.9682,遗传距离为0.0323。这些结果表明,紫色和茶绿色三疣梭子蟹群体的遗传多样性程度接近,具有相似的遗传多样性基础。另外,有2个随机引物（SBSA09和SBSG16）扩增得到群体特异性片段各1条。     

岱衢洋与官井洋大黄鱼养殖群体遗传多样性的AFLP分析

应用扩增片段长度多态性技术(AFLP),比较研究了岱衢洋大黄鱼养殖群体和官井洋大黄鱼养殖群体的遗传多样性。9对选择性引物组合,在2个群体的40个个体中共扩增出381个位点,其中多态位点为288个。2个养殖群体的多态位点比例、Nei′s基因多样性指数、Shannon多样性指数分别为65.01%和57.50%,0.162 3和0.1175,0.254 7和0.185 6,显示岱衢洋大黄鱼F2代养殖群体的遗传多样性高于官井洋大黄鱼繁育多代养殖群体的遗传多样性。UPGMA聚类分析图显示2个群体各自聚成一支。遗传分化系数Gst及AMOVA分析表明,2个群体的遗传变异主要来源于群体内个体间,但群体间已有一定程度的遗传分化。     

橙色莫桑比克罗非鱼和荷那龙罗非鱼的AFLP分析

运用AFLP分子标记技术检测了橙色莫桑比克罗非鱼（Oreochromis mossambicus）和荷那龙罗非鱼（Oreochromis hornorum）的种群遗传多样性。提取2种鱼的基因组DNA，经EcoR Ⅰ和Mse Ⅰ限制性内切酶酶切并加接头，从16对选扩引物组合中选出3对扩增结果较好的引物进行PCR扩增。扩增产物经PAGE胶电泳、银染，检测其多态性。3对引物在2种罗非鱼的40个个体中共扩增出94条带，其中多态性条带76条，占总扩增条带的80．9％，2个群体的多态性条带比例分别为71．6％和52．4％。根据Nei氏（1978）统计分析得出橙色莫桑比克罗非鱼种群内遗传相似系数为0．765。荷那龙罗非鱼的为0，821，表明2个罗非鱼种群均具有较高的遗传多样性，橙色莫桑比克罗非鱼种群内遗传多样性相对更高些。2种罗非鱼的种间遗传距离为0．279，推测该杂交组合可能具有杂种优势。     

管角螺遗传多样性的AFLP分析

近年来,由于对管角螺的捕捞强度加大,其自然资源大大减少。为进一步了解管角螺的遗传变异和群体结构状况,本文首次利用AFLP(Amplified fragment length polymorphism,扩增片段长度多态性)分子标记技术对我国南方沿海管角螺5个地理群体(连云港、温州、台山、湛江、防城港)进行遗传多样性分析。采用2对AFLP引物组合对管角螺5个群体的150个个体进行扩增,共扩增出310个位点,多态位点277个,多态位点比例为89.35%,基因多态性分别为0.382 4、0.476 6、0.393 8、0.437 4和0.392 8,说明5个群体的遗传多样性存在一定的差异。群体间及群体内遗传分化指数表明,管角螺5个地理群体的遗传变异主要来自于群体内个体间,而群体间无明显的遗传分化。UPGMA(非加权组对平均法)系统树分析表明,湛江和防城港群体间的亲缘关系较近,而台山和温州群体间的亲缘关系较远。     

虾类遗传多样性的研究现状

同工酶技术已经被广泛应用于虾类群体遗传学及标记辅助选择育种等研究方面.同工酶电泳显示对虾有相应的亚群差异,暖水性虾类的遗传多样性水平高于冷温性种类;连续分布的且在不同生活史阶段所处的栖息地环境变化幅度大的种类往往表现出较低的遗传多态性,虾类群体的多态位点比例和杂合度都较低,养殖群体比野生群体更低.但同工酶电泳技术往往检测不出一些"隐性"或"中性"变异而低估了遗传多样性水平.随着分子生物学技术的发展,从20世纪90年代开始,虾类遗传多样性的研究逐渐向线粒体DNA和核DNA的多态性方向发展.限制性片断长度多态性(RFLP)在虾类线粒体多倍型、物种标记以及比较野生和养殖群体线粒体COI 基因的多态性比较等方面应用普遍.随机扩增多态性(RAPD)技术主要应用于虾类不同地理群体遗传多态性调查,养殖群体与野生群体之间、养殖群体世代之间的遗传多态性比较,该技术得到的虾类遗传多样性水平高于同工酶电泳技术.扩增片断长度多态性(AFLP)技术在虾类中主要应用于遗传连锁图谱的构建,有关虾类的AFLP遗传多样性分析的报道不多.利用AFLP技术对连续选育群体的遗传多样性研究显示,随着选育世代的增加,选育群体的遗传多样性呈现下降趋势,但随着选育时间的延长,群体之间的分化逐渐降低,群体的遗传结构开始趋于稳定,成为一个品系.单一重复序列(SSR)技术主要用于标记种群遗传特异性、不同地理群体间以及野生和养殖群体间的遗传多样性差异.DNA序列分析技术特别是线粒体序列分析技术近年来在虾类遗传多样性研究中的应用逐渐增多.mtDNACOI、mtDNA12S rRNA和mtDNA16S rRNA的基因序列差异被应用于形态相近物种的鉴别、亚属分类、确定个体间亲缘关系和野生与养殖群体间的遗传多样性差异等.为了实现对虾类种群遗传差异的更深入了解,人们把种群遗传结构的研究和系统地理学相结合,即用分子系统地理学的方法来研究遗传谱系空间分布的历史特征,通过种群遗传结构的分析来探讨种内系统地理格局的形成机制、系统发育关系以及现有分布特征,并结合种群的地理分布状况来发现和验证与其相关的地质事件,追溯和揭示种群的进化历程.从育种、养殖以及资源保护的角度出发,人们开始研究野生群体的种质基因库,希望根据群体遗传结构确定合理的增殖策略,制定科学的保护措施,确保虾类养殖业的持续发展.     

条斑紫菜(Porphyra yezoensis)遗传多样性的AFLP分析

利用扩增片段长度多态性技术(AFLP)分析了13个条斑紫菜品系,在80对引物中,有11对能得到重复性好的多态性扩增条带,共扩增出619条谱带,每对引物扩增带数为40—77条,并且各对引物扩增结果均显示样品间存在差异。共得到609条多态性条带,多态位点的比例高达98.38%。根据AFLP图谱计算了不同样品间的遗传距离(GD)和相似性系数(GS)。聚类结果显示,品系海安与二室间的遗传相似系数较高而聚合在一起,不同样品间的相似性系数介于0.595—0.845之间。结果表明,条斑紫菜的遗传多样性极为丰富,而部分材料间的遗传距离有随地理间距增大而逐渐增大的趋势。聚类分析结果反映了目前条斑紫菜养殖品种间存在着相互混杂。     

军曹鱼的分子遗传特性研究

采用随机扩增多态性DNA(RAPD)和线粒体DNA(mtDNA)的限制性片段长度多态性(RFLP)分析方法对广东湛江近海军曹鱼Rachycentron canadum的分子遗传学特征进行了研究。RAPD研究结果显示17个引物共检测到119个位点,其中多态位点比例P为80．85％,遗传相似系数S为0．7400,遗传距离D为0．2600,Nei基因多样性指数H为0．3009,Shannon信息指数I为0．4498。结果表明,军曹鱼的遗传多样性比较丰富。用19种识别5、6碱基的限制性内切酶对军曹鱼的mtDNA RFLP进行了分析研究,构建其mtDNA物理图谱;用引物5’-GTG ATC TGA AAA ACC ACC GTT G-3’和5’-AAT AGG AAG TAT CAT TGC GGT TTG ATG-3’扩增线粒体细胞色素b区段,得到稳定的850bp大小的特异性条带,用18种识别4-6碱基的限制性内切酶对扩增片段的限制性长度多态性进行分析,并测定其序列。     

龙须菜RSAP分析及其SCAR标记的转化

初步探讨限制性位点扩增多态性(Restriction site amplified polymorphism,RSAP)分子标记技术在龙须菜遗传多样性检测和种质鉴定上的应用。利用所优化的适合于龙须菜RSAP分析的PCR反应体系,对青岛湛山湾野生群体和栽培品系981、07-2进行遗传多样性分析和比较。试验采用15对引物组合在3个群体14个龙须菜个体中共扩增出669个位点,其中多态位点数为146个,多态性比例22%,平均每对引物产生10条多态性条带,片段大小在100~1 000 bp之间。湛山湾野生群体的Na(1.007 5)、Ne(1.007 1)、H(0.003 6)、I(0.005 1)与栽培品系981的Na(1.003 0)、Ne(1.002 1)、H(0.001 2)、I(0.001 8),以及栽培品系07-2的Na(1.009 0)、Ne(1.006 3)、H(0.003 7)、I(0.005 4)相比较可知3个群体的遗传多样性是有差异的。种群间的遗传多样性分析表明,在所有检测的样品中,遗传多样性多来自不同群体间的多样性。群体遗传结构分析表明,2个栽培品系981、07-2间遗传距离不大,但栽培品系与湛山湾野生群体间的遗传距离相对较大,而UPGMA聚类分析也明显的将湛山湾野生群体与栽培品系981、07-2区分开来,表明野生群体与栽培品系间已产生一定的遗传隔离。通过分析湛山湾野生群体及栽培品系981、07-2的RSAP指纹图谱,从中筛选栽培品系981的特异条带,并将其转化为稳定性好、特异性高的序列特征化扩增区(Sequence characterized amplified regions,SCAR)分子标记。     

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.     

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。     

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.     

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.     

Deep-Sea Sediment Compression Curves: Some Controlling Factors, Spurious Overconsolidation, Predictions, and Geophysical Reproduction

Oedometer tests have been carried out on 70 undisturbed surficial clays (at approximately 250 mm below the mudline), mostly collected by free-fall corers from sites widely scattered throughout the deep-sea North Atlantic. Acoustic measurements were also made, initially on contiguous samples and ultimately on the same sample using a geophysically instrumented oedometer which also collected electrical resistivity data. Apart from those quiescent areas below the carbonate compensation depth, such as north of the West Indies where very fine clays exist, most of the samples are silty clays whose geotechnical-geophysical properties are dependent on the type of clay minerals present (and their ability to take in moisture), the sand-size fraction, and the quantity of carbonate present. Thus the pure clays have high compressibilities which decrease on the addition of coarse particles, while the converse is true for the acoustic parameters, these increasing with the sand fraction. Using the notion of the intrinsic compression line for all samples, and comparison to it of the measured compression curves, it is clear that, contrary to some previously held ideas, most deep-sea clays are normally consolidated; the addition of carbonate has the effect of creating an open, stronger sediment skeleton. Interestingly, where information is available, the variation with depth of a sample's acoustic velocity follows the void ratio pressure relationship of the compression curve. This allows the construction of an in-situ sediment compression curve using the in-situ geophysical observations.     

Phylogenetic diversity and phenotypic characterization of cultivable bacterioplankton isolated from polar oceans

A set of 27 marine planktonic bacteria isolated from the polar regions was characterized by 16S rDNA sequencing and physiological and biochemical testing. More than half of these bacteria were positive for caseinase, gelatinase and 13-glucosidase, and could utilize glucose, maltose or malic acid as carbon source for cell growth. Twelve isolates expressed nitrate reduction activities. Except for one antarctic isolate BSwlO175 belonging to Actinobacteria phylum, these isolates were classified as γ-Proteobacteria, suggesting that γ-Proteobacteria dominated in cultivable marine bacterioplankton at both poles. Genus Pseudoalteromonas was the predominant group in the Chukchi Sea and the Bering Sea, and genus ShewaneUa dominated in cultivable bacterioplankton in the Prydz Bay. With sequence similarities above 97%, genus Psychrobacter was found at both poles. These 27 isolates were psychrotolerant, and significant 16S rDNA sequence similarities were found not only between arctic and antarctic marine bacteria （ 〉 99% ）, but also between polar marine bacteria and bacteria from other aquatic environments （ ≥ 98.8% ）, including temperate ocean, deep sea, pond and lake, suggesting that in the polar oceans less temperature-sensitive bacteria may be cosmopolitan and have a bipolar, even global, distribution at the species level.     

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.