真鲷三倍体诱导初步研究

在南鲷Ｐａｇｒｏｓｏｍｕｓ ｍａｊｏｒ三倍体诱导研究中，对染色体倍数的分析表明，受精卵受精５ｍｉｎ后，在１．５℃冰水中处理５ｍｉｎ可获得１７％的三倍体诱导率和２８％的嵌合体比率，染色体数分别是：２Ｎ＝４８，３Ｎ＝７２，嵌合体５０－６８。仔鱼红血细胞测量和组织切片分析显示，三倍体和二倍体的红血细胞核直径，视网膜神经节细胞核直径物中脑细胞核直径大小存在差异，其比率分别是１．４２，１．３５和１．４１，这     

半滑舌鳎三倍体诱导研究

采用冷休克抑制第二极体排出的方法诱导了半滑舌鳎三倍体,并探讨了三倍体诱导的最佳条件.通过流式细胞仪倍性检测和染色体观察验证各组三倍化率.结果显示:在受精后5 min开始,将受精卵放在4℃海水中进行冷休克处理,持续时间为20 min,三倍体诱导率最高为35％.通过对三倍体染色体进行分析,辨认其雌雄染色体型为ZWW/ZZZ,出现频率为1∶1,没有发现ZZW型雌鱼,暗示半滑舌鳎Z染色体和W染色体之间的交叉和互换受到严重抑制.     

杂交三倍体太平洋牡蛎群体的染色体数目组成初步观察

探讨多倍体牡蛎的遗传基础,通过染色体计数法对2龄杂交三倍体太平洋牡蛎(Crassostrea gigas)群体的染色体数目组成进行了分析,发现在80个具良好中期分裂相的牡蛎中,有56个三倍体(70.0%);12个非整倍体(15.0%),4个2n/3n嵌合体(5.0%);8个二倍体(10.0%)。12个非整倍体中包括9个29条染色体的亚三倍体(3n-1)和3个31条染色体的超三倍体(3n 1)。非整倍体与三倍体的生长差异不明显(P>0.05)。可能由于数量较少的原因,嵌合体和二倍体的生长没有表现明显生长滞后(P>0.05)。     

三倍体皱纹盘鲍(Haliotis discus Hannai)性腺发育的生物学研究

采用光镜和电镜方法，观察了吊笼式人工养殖三倍体皱纹盘鲍性腺发育情况，并与二倍体进行比较。结果表明，三倍体鲍性腺发育分期基本与二倍体相同，但雄性腺发育停止在生长期，雌性腺少数可达成熟期，没有发现排放期。三倍体鲍性腺发育较二倍体差，表观看其性腺薄，只包围肝胰腺1／3—1／2，颜色浅：7—10月间检查的86个二倍体个体中成熟期的占24．4％，休止期占11．6％。而83个三倍体个体中，成熟期占1．2％，休止期占72．3％。三倍体成熟卵母细胞直径、核径、线粒体直径、胶膜厚度较二倍体大，但卵黄颗粒、线粒体、粗面内质网等细胞器较二倍体少。     

华贵栉孔扇贝的三倍体诱导及生长比较

分别用细胞松弛素Ｂ（ＣＢ，浓度０．５ｍｇ·Ｌ（－１））和低温（１０℃）处理诱导华贵栉孔扇贝Ｃｈｌａｍｙｓｎｏｂｉｌｉｓ产生三倍体。抑制第一极体的三倍体诱导率分别达到４４．１％和３１．８％；而抑制第二极体的三倍体诱导率分别为９０．２％和７２．７％（胚胎初期检查）。ＣＢ处理组幼虫的日增长率显著超过二倍体（ｐ＜０．０１）。养殖半年后，三倍体在壳高、壳长、壳宽和体重方面均显著大于二倍体（ｐ＜０．０１）；而养殖１３个月后，三倍体的肉重和闭壳肌重分别比二倍体增加３９．５％和６７．４％。在繁殖期，三倍体在外观上没有生殖腺。     

大黄鱼倍性检测研究

于2001—2004年对人工诱导大黄鱼的不同发育阶段和不同组织进行倍性检测，胚胎眼泡期采用染色体计数法测定，鱼苗和成鱼阶段用流式细胞法，结果表明，胚胎染色体数目2n=48条为二倍体，3n=72条为三倍体．采用流式细胞法测定鱼苗和成鱼个体时，若受测样品的DNA相对含量值与已知对照组二倍体的比值约为1．5：1，其个体为三倍体；比值约为1：1，则为二倍体，无论用肌肉或尾鳍组织，皆可检测出其个体的倍性。     

不同倍性大黄鱼核仁数目银染观察

用银染法对不同倍性(单倍体、正常二倍体、正常三倍体、雌核发育二倍体、雌核发育鱼子代三倍体)大黄鱼胚胎、仔鱼或成鱼的不同组织进行细胞核仁数目观察计数。结果表明,大黄鱼各种倍性个体细胞核仁数目与染色体倍性之间有良好的对应关系,且核仁数随个体倍性的增加而增加;非诱导雌核发育的大黄鱼细胞核仁众数与染色体组数相等(二倍体为2,三倍体为3),而人工诱导雌核发育大黄鱼及其子代细胞核仁众数却比其相同倍性的非雌核发育鱼少1(雌核发育二倍体为1,雌核发育三倍体为2)。雌核发育鱼核仁数减少是一个非常有趣的现象,值得进一步研究。大黄鱼不同发育阶段(胚胎、仔鱼、成鱼)以及同一个体不同组织(鳃、肾、鳍)的细胞核仁数目(众数)相同。因此,借助核仁银染法可以快速、可靠、简便地进行染色体倍性鉴定。而雌核发育鱼核仁数减少的机理值得进一步研究。     

三倍体和二倍体合浦珠母贝育珠比较初步研究

利用人工诱导培育的三倍体合浦珠母贝Pinctada martensii(D.)进行插核育珠试验,并同处理组内的二倍体同胞进行比较。结果表明:三倍体合浦珠母贝的留核率和正圆珠比率分别为81.0％和58.8％,而二倍体则为71.3％和37.8％;在非正圆珠中两者的污珠率差别不大,但三倍体的尾巴珠和素珠的比率明显低于二倍体;三倍体的正圆珠珍珠层平均厚度和平均重量分别为381μm和163mg,二倍体分别为265μm和105mg,两者差异极显著(P<0.01)。初步表明,利用三倍体合浦珠母贝培育珍珠,能缩短育珠期,提高珍珠的产量和质量。     

二倍体和三倍体太平洋牡蛎(Crassostrea gigas)繁殖前后生化成分及氨基酸组成的比较研究

对太平洋蛎二倍体和三倍体在繁殖前后的几种生化成分和氨基酸组成进行了比较研究，为牡蛎的生理生化研究提供了基础数据。结果显示：太平洋牡蛎的总糖，糖原，脂肪和蛋白的含量与其繁密切相关，二倍体牡蛎在繁殖后的总糖，糖原，脂肪的含量比繁殖前分别降低了24．31％，30．37％，14．91％，三倍体分别降低了9．45％，7．17％，20．29％，而二倍体和三倍体牡蛎粗蛋白的含量在繁殖后较繁殖前分别增高了14．91％，12．285，表明了糖类和脂肪是牡蛎殖能量的主要来源，繁殖前三倍体牡蛎的糖原含量比二倍体高23．30％，繁殖后二倍体糖原含量明显降低，三倍体的糖原含量比二倍体高了73．83％，表明三倍体牡蛎由于可育性低而体内一直保持较高水平的糖原含量，从而在繁殖季节也保持了鲜美的口味。二倍体太平洋牡蛎与三倍体的氨基酸组成和含量的分析比较也表明，二倍体牡蛎的氨基酸组分在繁殖前后变化较大，而三倍体的氨基酸组分相对稳定。     

抑制栉孔扇贝第一极体对受精卵染色体行为及胚胎倍性组成的影响

采用细胞松弛素B（CB）处理，栉孔扇贝（Chlamys ferreri）抑制其受精卵的第一极体（PB1），研究抑制PB1对受精卵减数分裂过程及胚胎倍性组成的影响。结果发现，抑制第一极体显著改变了受精卵的染色体行为，在第二次减数分裂过程中共发现4种典型染色体分离类型，分别是三极分离（41．7％）、二极分离（11．7％）、双二极分离（24．9％）和非同步分离（2．8％），其余的受精卵（19．0％）染色体分离行为紊乱。对4-8细胞期胚胎的倍性组成进行分析，发现处理组中含有二倍体（10．9％）、三倍体（12．5％）、四倍体（19.5％）、五倍体（12．6％）以及非整倍体（46．6％）胚胎。研究结果表明，二极分离和双二极分离分别是形成三倍体和四倍体的主要机制，而其他的染色体分离行为将主要形成非整倍体。     

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.