盐藻分子生物学研究概况

杜氏藻(Dunaliella sp．)又称盐藻,是一类极端耐盐的单细胞真核绿藻,属于真绿藻纲、杜氏藻目、杜氏藻科、杜氏藻属．有些杜氏藻能够耐受环境中NaCl浓度在0．05％～39．4％之间的变化,可以在pH5．5～9．5的范围内生存。杜氏藻细胞体内能储存大量的甘油和胡萝卜素等有机化合物。可通过大量培养杜氏藻来提取β-胡萝卜素。天然甘油是良好的化妆品基质。而β-胡萝卜素是食品添加剂和着色剂。且具有医疗保健功能;     

盐藻生长过程中氮磷利用与色素积累

考察了盐藻在氮浓度3mmol/L和0.59mmol/L两种条件下的生长、氮磷的利用以及色素积累的情况。结果表明氮浓度3mmol/L培养基有利于藻细胞的增值，最终藻体浓度大，稳定期可维持数日；0．59mmol/L培养基有利于细胞胡萝卜素积累，单位细胞胡萝卜素与叶绿素的比值达到12．3，是氮浓度3mmol/L培养基的两倍左右。盐藻细胞在生长初期快速利用氮源，之后即使培养基中仍有较高浓度的氮源，藻细胞也不再利用。两种条件下藻细胞对磷的利用情况相同。两种培养基中盐藻单位细胞叶绿素含量均先增加后减少，胡萝卜素含量均先降低后增加。     

光照、温度和营养盐对三株盐生杜氏藻生长和色素积累的影响

通过正交试验研究了光照、温度和营养盐对3株盐生杜氏藻株系(BJ,OUC38和OUC8)生长和色素积累的影响,旨在探明适于盐生杜氏藻生长和β-胡萝卜素积累的光照、温度和营养盐条件,并对实验中3株株系生长和β-胡萝卜素积累速率进行了初步的比较。结果表明,盐生杜氏藻生长的最适光照和温度分别是145.3μmol.m-2.s-1和23℃;适于β-胡萝卜素积累的光照和温度分别是218.2μmol.m-2.s-1和23℃;适于生长的营养盐条件是NaNO30.4 mmo/L、尿素0.6 mmol/L和NaHCO318 mmol/L,而当NaNO3、尿素和NaHCO3浓度分别为0,0.2和18 mmol/L时有利于β-胡萝卜素的积累。高光照、低温有利于杜氏藻细胞β-胡萝卜素的积累,在218.2μmol.m-2.s-1光强和15℃条件下藻细胞β-胡萝卜素积累平均值可分别达10.39和12.46 pg/cell。3个株系中OUC38积累β-胡萝卜素的能力高于另外2株,但差异不显著。     

培养密度和培养液“老化”对盐藻生长和β-胡萝卜素累积的影响

通过研究培养密度和培养液“老化”对盐生杜氏藻Ｄｕｎａｌｉｅｌｌａ ｓａｌｉｎａｗ Ｔｅｏｄｏｒｅｓｃｅ生长和β－胡萝卜素累积的影响，发现盐藻的细胞增长率在接种后第２天达到高峰，然后逐渐下降；β－胡萝卜素的日累积量在接种后逐渐增加，第７天达到高峰，然后下降，这一现象与在培养过程中细胞密度上升而使每个细胞所接收的光强下降以及培养液的“老化”有关。     

盐藻SOD的盐适应性与物质积累的关系

实验分析了不同NaCl浓度处理下,盐藻SOD活性及其与细胞密度、β-胡萝卜素积累和蛋白质积累的关系.结果表明:在30～120 g/L的盐度范围内,随着NaCl浓度的提高,SOD活性极显著地增强,说明盐藻SOD是一种高盐适应酶.盐藻SOD活性与细胞密度及物质积累关系密切:在SOD活性为0.710×108 cells-1·min-1左右时,盐藻细胞密度、β-胡萝卜素和蛋白质的积累量都最高,而在SOD活性较低或较高时,盐藻细胞密度、β-胡萝卜素和蛋白质的积累量都较低.可能适当盐度下SOD的活动较好地保护了盐藻体内物质积累反应的进行.     

盐藻过氧化物酶的盐适应性及其与物质积累的关系

实验分析了不同浓度NaCl处理下,培养盐藻的过氧化物酶(POD)活性及其与细胞密度、β-胡萝卜素积累和蛋白质积累的关系.结果表明,盐藻过氧化物酶活性随盐度变化而改变:在适当盐度(60～90g/L)下,过氧化物酶活性很低;在较低盐度(30～60g/L)或较高盐度(90～150g/L)下,盐藻过氧化物酶活性均显著升高,说明盐藻过氧化物酶是一种盐度逆境适应酶.盐藻过氧化物酶活性与盐藻细胞密度及物质积累关系密切:盐藻过氧化物酶活性很低时,盐藻细胞密度大,同时β-胡萝卜素和蛋白质积累也多;随着盐藻过氧化物酶活性升高,盐藻细胞密度、β-胡萝卜素和蛋白质积累均逐渐降低,但盐藻过氧化物酶活性进一步升高时,盐藻蛋白质积累又有增加,可能在盐藻体内有逆境蛋白产生.     

胁迫条件下盐藻β-胡萝卜素及其异构体累积的研究──盐度的影响

于１９８９－１９９１年，在青岛室外对盐藻为实验材料，利用Ｊｏｈｎｓｏｎ改良配方制作增养液，进行盐度（９０－２４０）实验，结果表明，细胞生长的适宜盐度为１２０；当盐度超过１２０－２４０，盐度越高对细胞生长越不利，但越有利于细胞内β－胡罗卜素的累积；适宜于养殖盐藻生产β－胡萝卜素中至少含６种异构体，盐度增加不利于顺式异构体的累积。为生产防癌和延缓衰老等作用更强的顺式异构体，建议生产性养殖时适当降低盐度     

氮磷比对盐藻生长及甘油和色素累积的影响

将盐藻在［Ｎ］／［Ｐ］（氮磷浓度比）为０．５至６０．０的培养液中进行培养,测定了不同实验条件下［Ｎ］／［Ｐ］比值对盐藻生长、培养液中甘油和叶绿素含量以及β－胡萝卜素累积的影响,并初步建立了盐藻在不同［Ｎ］／［Ｐ］比值下的生长动力学模式。研究结果表明,在［Ｎ］／［Ｐ］为２５时,盐藻的生长最佳。［Ｎ］／［Ｐ］比值变化对甘油含量的影响不大。［Ｎ］／［Ｐ］比值小于１５时,叶绿素含量随［Ｎ］／［Ｐ］比值的增加而增加;比值大于１５时,叶绿素含量基本不受［Ｎ］／［Ｐ］比值变化的影响．单细胞β－胡萝卜素含量随［Ｎ］／［Ｐ］比值的增加而减少,而培养液中β－胡萝卜素含量在［Ｎ］／［Ｐ］比值小于１５时基本不受影响,比值大于１５时含量呈下降趋势。     

杜氏藻的RF-RAPD分子系统学分析

杜氏藻(Dunaliella)是绿藻门、绿藻纲、团藻目、多毛藻科中的一个属(也有单独归为杜氏藻目、杜氏藻科、杜氏藻属[1]).因可以在高盐的极端环境中生存,也称之为盐藻.盐藻富含各种类胡萝卜素和甘油,其中β-胡萝卜素累积可达细胞干重的14%,具有较高的经济价值.另外盐藻无细胞壁,仅有一层糖蛋白和神经氨酸组成的外膜包裹,类似天然的原生质体,是真核细胞基因工程首选的受体之一.盐藻主要用来生产β-胡萝卜素,生产中发现β-胡萝卜素产量不稳定,推测种质差异因素与此有密切关系,因此分析不同种及种内品系及其与β-胡萝卜素含量相关性非常重要.     

不同氮源对盐生杜氏藻生长和色素积累的影响

研究了NaNO3、NH4Cl、NH4NO33种氮源对盐生杜氏藻(Dunaliella salina)生长和色素积累的影响及3种氮源吸收的规律。结果表明,3种氮源中,NH4NO3对促进D. salina细胞生长、β-胡萝卜素积累和叶绿素a合成效果最好,细胞最高密度为136×104cell/mL;β-胡萝卜素含量最大值为137mg/g;NO3-和NH4 共同存在时,优先利用NH4 ;建立了3种氮源吸收的动力学方程。     

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。     

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.     

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.     

INFORMATION FOR AUTHORS

<正>Acta Oceanologica Sinica(AOS)is a comprehensive academic journal edited by the Editorial Committee of Acta Oceanologica Sinica and is designed to provide a forum for important research papers of the marine scientific community which reflect the information on a worldwide basis.The journal publishes scholarly papers on marine science and technology,including physics,chemistry,biology,     

Estimation of Sediment Properties during Benthic Impact Experiments

Sediment properties, such as water content and density, have been used to estimate the dry and wet weights, as well as the volume of sediment recovered and discharged, during benthic impact experiments conducted in the Pacific and Indian Oceans. The estimates show that the weights of dry (355-1332 t) and wet sediment (1651-4888 t) recovered during the experiments, are not only a function of the total duration and distance covered during the experiment, but also depend on water content and density of the sediment. Estimation of dry sediment and its volumetric ratio in the discharge, are key parameters for calculating the discharged volume. The estimated volume of wet sediment recovered (1427-4049 m 3 ) as well as discharged (2693-6951 m 3 ) during the experiments, provide important inputs to evaluate the sediment resettlement and migration. Using these estimates, the average depth of excavation on the seafloor, can also been calculated. However, the stages of sediment recovery and discharge are expressed differently in some of the experiments. In order to standardize these, different stages have been identified, and definitions of certain terms have been suggested, for use in the future. The methods of calculating different properties, as well as weight and volume of discharged sediment are described in the paper for use in other applications concerning deep-sea discharges.