一种新方法——低渗诱导虾夷扇贝三倍体的研究

研究开发了1种多倍体诱导的新方法,采用低渗抑制受精卵第2极体(PB2)的释放诱导虾夷扇贝(Patinopecten yessoensis)三倍体.结果表明:以低渗盐度S=12,在第一个PB2出现时开始处理,持续处理20 min,诱导率可高达(84.74±3.82)%;三倍体处理组的幼虫比对照组具有明显的生长优势,但是幼虫存活率低于二倍体对照组.     

合浦珠母贝人工诱导四倍体的研究

用细胞松驰素B（CB）、聚乙二醇（PEG）和静水压抑合浦珠母贝Pinctadamarlensii（D．）制第1次卵裂以及CB抑制极体形成诱导四倍体。CB抑制第1次卵裂在早期胚胎2-4细胞阶段发现四倍体,但8细胞阶段以后四倍体胚胎消失。PEG和PEG＋CB能诱导细胞融合产生四倍体,但处理组幼虫在担轮期死亡。CB抑制极体形成能诱导出较高比例的四倍体,在胚胎初期和担轮幼虫期分别占40％和30％以上。处理组幼虫进行培育,附苗后当贝苗长成4-5cm大小时通过鳃细胞染色体倍性检查,却没有发现四倍体。文中对四倍体是否能成活进行了讨论。     

利用细胞松弛素B抑制第一极体排放诱导近江牡蛎(Crassostrea rivularis)四倍体

采用细胞松弛素B（CB）处理受精卵,抑制第一极体的排放,对近江牡蛎四倍体的诱导和培育进行了研究.结果表明,近江牡蛎在25-28℃时受精卵发育同步性较高.在28℃条件下,CB适宜处理剂量为0.6mg/L.在个别受精卵出现第一极体时实施处理,至对照组的第一极体出现率达到50%终止处理,四倍体的诱导效果较好.5个处理组D形幼虫的平均孵化率为11.0%,显著低于对照组72.4%的平均孵化率.在受精后4-12d期间处理组幼虫的存活率显著低于对照组.处理组胚胎和D形幼虫阶段四倍体的平均比例分别为41.8%和37.9%.四倍体的比例随着幼虫的生长发育呈下降趋势,至受精后12d在各处理组已检测不到四倍体.2组幼虫完成附着变态,从受精卵至稚贝的累计存活率平均为0.3%.本研究结果表明,使用CB处理抑制受精卵第一极体的释放可以有效诱导近江牡蛎四倍体,但所获得的四倍体幼虫存活力差,需要在未来的研究中解决.     

栉孔扇贝第1卵裂抑制型雌核发育二倍体的研究

利用6-二甲基氨基嘌呤(60μg/cm3;6-DMAP)和细胞松弛素B(0.5μg/cm3;CB)抑制受精卵的第1次卵裂,授精后60和65 min分别用CB和6-DMAP持续处理20和15 min,可诱导出12.5%和27.5%的第1卵裂抑制型雌核发育二倍体.细胞学观察显示,6-DMAP抑制第1次卵裂产生的雌核发育二倍体主要在第1次有丝分裂后期的受精卵,由于破坏了纺锤体和阻止了染色体分离,导致一个二倍性雌核的形成,而CB有效地阻止了第1次卵裂的胞质分裂.尽管倍化率和D型幼虫发生率较低,但该研究首次报道了栉孔扇贝第1卵裂抑制型雌核发育二倍体诱导的可行性.     

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。     

栉孔扇贝四倍体的研究Ⅰ:细胞松驰素B诱导

利用细胞松驰素Ｂ抑制栉孔扇贝Ｃｈｌａｍｙｓ（Ａｚｕｍａｐｅｃｔｅｎ）ｆａｒｒｅｒｉ受精卵的第一极体的放出和第一次有丝分裂获得四倍体。在２０℃条件下，卵子受精后１０ｍｉｎ，０．５μｇ／ｍｌ的ＣＢ持续处理１０ｍｉｎ、２０ｍｉｎ抑制第一极体排出，结果三倍体率和四倍体率分别为３８．３８％、３９．７５％和２８．４２％、３０．２４％；在２０℃条件下，卵子受精后１ｈ，０．５μｇ／ｍｌ的ＣＢ持续处理１０、１５、２０、２５ｍｉｎ抑制第一次有丝分裂，结果四倍体率分别为２１．３６％、２７．５７％、２８．４１％、２９．３５％。结合处理后幼虫的Ｄ形率，在抑制第一极体放出和第一次卵裂中，ＣＢ处理持续１０ｍｉｎ均为最好     

用6—甲氨基嘌呤诱导贻贝四倍体胚胎

在滤过海水中以４×１０＾－４ｍｏｌ·Ｌ＾－１６－甲氨基Ｃ嘌呤（６－ＤＭＡＰ）处理贻贝Ｍｙｔｉｌｕｓ ｅｄｕｌｉｓ受精卵或胚胎，受精激活后５０或１２０ｍｉｎ处理２０－３０ｍｉｎ，分别抑制第一或第二次有丝分裂，以诱导四倍体胚胎。顶荧光显微观察表明，６－ＤＭＡＰ有效地抑制了第一和第二次卵裂。它使细胞核染色质分散，抑制了原核的移动和染色体的分离，并防碍卵裂沟和极叶的形成，从而诱导出四倍体胚胎。对第一次卵裂     

合浦珠母贝人工诱导四倍体的研究

用细胞松驰素Ｂ（ＣＢ）、聚乙二醇（ＰＥＧ）和静水压抑合浦珠母贝Ｐｉｎｃｔａｄａ ｍａｒｔｅｎｓｉｉ（Ｄ．）制第１次卵裂以及ＣＢ抑制极体形成诱导四倍体。ＣＢ抑制第１次卵裂在早期胚胎２－４细胞阶段发现四倍体,但８细胞阶段以后四倍体胚胎消失。ＰＥＧ和ＰＥＧ＋ＣＢ能诱导细胞融合产生四倍体,但处理组幼虫在担轮期死亡。ＣＢ抑制极体形成能诱导出较高比例的四倍体,在胚胎初期和担轮幼虫期分别占４０％和３０％以上。处     

栉孔扇贝四倍体诱导效果的早期分析方法

应用流式细胞仪检测栉孔扇贝(Chlamys farreri)担轮幼虫、D型幼虫倍性组成，结合现场观察、压片观察及D型幼虫生成率分析了热休克抑制第一次卵裂(MI)诱导栉孔扇贝四倍体的诱导效果。该方法可快速、有效地分析出处理剂量、起始处理时间、处理持续时间对诱导效果的影响。从而筛选出合适的诱导指标。     

用6－甲氨基嘌呤诱导贻贝四倍体胚胎

在滤过海水中以４×１０（－４）ｍｏｌ·Ｌ（－１）６－甲氨基Ｃ嘌呤（６－ＤＭＡＰ）处理贻贝Ｍｙｔｉｌｕｓｅｄｕｌｉｓ受精卵或胚胎，受精激活后５０或１２０ｍｉｎ处理２０—３０ｍｉｎ，分别抑制第一或第二次有丝分裂，以诱导四倍体胚胎。顶荧光显微观察表明，６－ＤＭＡＰ有效地抑制了第一和第二次卵裂。它使细胞核染色质分散，抑制了原核的移动和染色体的分离，并防碍卵裂沟和极叶的形成，从而诱导出四倍体胚胎。对第一次卵裂的抑制产生８２．８％的四倍体胚胎；第二次卵裂抑制产生５８．６％的四倍体胚胎。由于６－ＤＭＡＰ如此有效，而且价格便宜以及对人体无害，与沿用的化学诱导剂细胞松弛素Ｃ．Ｂ．相比，它应该可以成为双壳类软体动物染色体操作研究的理想化学诱导剂之一。     

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.     

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.     

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.     

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.     

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.     

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.