盐胁迫对盐生杜氏藻生长及叶绿素荧光特性的影响3期王帅,等:盐胁迫对盐生杜氏藻生长及叶绿素荧光特性的影响35

以盐生杜氏藻(Dunaliella salina)为实验材料,研究在不同盐度(5,10,20,30,40,50,60,70,80,90,100,110,120,130)下处理不同时间后(12、24和48h),该藻的生长及叶绿素荧光特性的变化情况。结果表明,盐生杜氏藻生长和进行光合作用的最适盐度为100,适宜盐度范围为20~110。在盐度过低(20)和过高(110)的胁迫条件下,盐生杜氏藻PSII的荧光指标(Fv/Fm、ΦPSII、Fv/Fo和rETR)显著降低。该藻的细胞密度和叶绿素含量均在盐度100时达到最大值。相关性分析结果表明,盐生杜氏藻的荧光指标(Fv/Fm、ΦPSII、Fv/Fo和rETR)与细胞密度呈极显著的正相关性,因此可通过PSII荧光指标的变化来反映该藻对盐胁迫的耐受性。本文还对盐胁迫下盐生杜氏藻的响应机制和叶绿素荧光技术在筛选耐盐微藻品种中的应用进行了初步探讨。     

盐度和光照强度对赤潮异弯藻增长的效应

采用一次性培养的方法,研究了盐度和光照强度对赤潮异弯藻增长的效应。结果表明:赤潮异弯藻在3种光照和7个盐度梯度下均可生长。盐度对赤潮异弯藻的生长有极显著的影响(P0.01),光照强度对赤潮异弯藻的生长影响亦显著(P0.05),盐度和光照强度的交互作用无显著影响(P0.05)。不论在哪种光照下,低盐度10、15组赤潮异弯藻虽然能生长,但生长速率和最大细胞数量值均较低,随着盐度的提高,藻的生长速率和最大细胞数量值增大,但盐度增至30以后,即30~40试验组的生长速率和最大细胞数量值之间的差异不明显。而不论在哪个盐度条件下,赤潮异弯藻在3种光照强度下,均以6000lx时的藻细胞密度值最高,生长情况最好。     

不同氮浓度下盐胁迫对坛紫菜(Pyropia haitanensis)生长和光合作用的影响

为了探讨在不同氮浓度条件下盐度胁迫对坛紫菜(Pyropia haitanensis)生长和光合生理的影响,设置高(500μmol/L)、低(50μmol/L)2个氮浓度和15、30和45三个盐度水平,将坛紫菜叶状体在不同的氮和盐度条件下适应培养7d后,测定藻体的生长、色素含量、光合放氧和快速光反应曲线等生理指标。结果显示:在低氮条件下,坛紫菜的生长速率随盐度增加而递减;在高氮下,30盐度水平培养的藻体具有最大生长速率。低氮条件下,叶绿素a(Chl a)、类胡萝卜素(Car)、藻红蛋白(PE)和藻蓝蛋白(PC)的含量在盐度15中较高,而在盐度30和45之间没有显著差异;高氮条件下,盐度对藻体Chl a和Car的含量没有显著影响,但相对于盐度30的处理,盐度15和45都显著降低了PE和PC的含量,虽然这两种色素含量在两个盐度处理间没有显著差异。不同的氮和盐度处理对藻体的光合放氧速率几乎没有影响,除了高氮条件下,单位湿重的放氧速率在盐度30中较低而单位Chl a的放氧速率在盐度45中稍高。从快速光反应曲线的结果看出,盐度虽然对光饱和点(Ik)、电子传递效率(α)和最大相对电子传递速率(r ETRmax)没有显著影响,但高盐胁迫显著提高了光抑制项的值(a)。综上所述,高盐度胁迫能够抑制坛紫菜的生长和光合作用,氮的浓度对这种抑制作用没有显著影响;低盐度胁迫对藻体的生长和光合没有抑制,甚至在低氮条件下表现出对生长的促进。这些结果为坛紫菜的栽培和对富营养化海水的修复提供了一定的理论参考。     

盐度胁迫对大海马(Hippocampus kuda)幼体生长、组分及酶活力的影响

在盐度为25的条件下,以大海马幼体为实验材料,通过设置不同的盐度胁迫组(盐度从25胁迫至5、15和35)的方法,对其生长、生化组分以及酶活力的影响进行了研究。结果表明:15盐度组大海马幼体的体重、生化组分、能值与对照组(盐度25)相比差异不显著(P>0.05),体长、成活率指标则显著高于对照组(P<0.05),然而5、35盐度组的生长指标、成活率、生化组分等则显著低于对照组(P<0.05)。15盐度组的SOD、CAT酶活性低于对照组水平(P<0.05),MDA的含量变化不显著(P>0.05);而5、35盐度组SOD、CAT和MDA含量与对照组相比,随着时间的延长,呈现逐渐升高的趋势(P<0.05);随着盐度的升高,AKP酶活性具有逐渐升高的趋势,而ACP酶活性则呈现降低趋势。     

盐度胁迫下中肋骨条藻和东海原甲藻的生长及内源多胺含量的变化

在实验室内不同盐度梯度下培养常见的两种赤潮藻——中肋骨条藻(Skeletonema costatum s.l.)和东海原甲藻(Prorocentrum donghaiense),根据logistic生长模型获得了终止生物量Bf和最大生长速率μmax,并测定了藻体中过氧化物酶(POD)、二胺氧化酶(DAO)、多胺氧化酶(PAO)的活性和丙二醛(MDA)、腐胺(Put)、亚精胺(Spd)、精胺(Spm)的含量。结果表明,高、低盐度胁迫下,两种藻均会积累MDA,活性氧自由基的伤害增强,藻的生长受到了抑制。同时,两种藻都会提升多胺含量,通过多胺的调节作用来缓解胁迫的伤害,促进生长,但是在提升的多胺种类和形态上,两种藻存在着差异。高盐胁迫下,中肋骨条藻会提升多胺尤其是游离态多胺的含量来缓解伤害,东海原甲藻则依靠结合态的亚精胺和游离态的精胺的调节作用。低盐胁迫下,中肋骨条藻会提升游离态腐胺的含量,而东海原甲藻体内各形态的多胺都会上升。     

温、盐度及营养盐对不同藻株塔玛亚历山大藻生长的影响

利用2001年11月至2003年2月从长江口、闽江口和北部湾水域表层沉积物中分离的休眠孢囊,萌发获得三株塔玛亚历山大藻,并研究了温度、盐度、营养盐对它们生长的影响。实验结果表明,来源于三个不同海区的藻株在水温32℃的条件下均无法存活,温度对长江口藻株(ATDH01)及广西北部湾藻株(ATGX02)的生长有极显著影响(P<0.01),而对闽江口藻株(ATMJ02)的生长无显著影响(P>0.05)。不同海区的藻株,其生长特性有明显差异,ATDH01藻株最适生长温度为24℃,最适生长盐度为30,且在盐度15和35仍有较高的生长率;而ATMJ02藻株及ATGX02藻株最适生长温度分别为24℃和20℃,最适生长盐度范围为20～30,盐度超出此范围生长率明显降低。ATDH01藻株的生长繁殖能力最强。营养盐对三个不同海区藻株生长的影响基本一致,在低P(1.2μmol/L)组中,三株藻的指数生长期较短,细胞密度明显的低于f/2组;而在低N(44μmol/L)组中,虽然在培养初期,三株藻的细胞生长明显快于f/2组,但在生长后期由于氮的限制,最终细胞密度仍低于f/2组。     

盐度胁迫下盐生隐杆藻抗氧化防御系统的变化

盐生隐杆藻(Aphanothece halophytica)是在饱和NaCl条件下可以生存的少数几种耐盐蓝藻之一,作者研究了盐胁迫对盐生隐杆藻细胞内丙二醛含量、2种非酶抗氧化剂含量以及4种抗氧化酶活性的影响。与对照组相比,低盐胁迫和高盐胁迫都抑制盐生隐杆藻的生长。随着盐度胁迫程度的增加,丙二醛含量、非酶抗氧化剂含量和抗氧化酶(除了过氧化氢酶)活性不断增强。上述结果表明:盐胁迫下,盐生隐杆藻细胞内非酶抗氧化剂和抗氧化物酶水平的增加可以应对细胞内升高的氧化伤害,抗氧化防御系统可能在盐生隐杆藻对盐胁迫的耐受机制中发挥了重要作用。     

盐胁迫对塔胞藻生长及叶绿素荧光动力学的影响

利用调制式叶绿素荧光仪(Water-PAM)研究了盐度(1~120)胁迫不同时间(12,24,48 h)后,塔胞藻生长及叶绿素荧光动力学的变化。测定的主要参数有:PSⅡ的最大光能转化效率(Fv/Fm)、PSⅡ的潜在活性(Fv/Fo)、PSⅡ的实际光能转化效率(ΦPSⅡ=yield)、光合电子传递效率(ETR)、光化学淬灭(qP)和非光化学淬灭(NPQ)。结果表明:在本实验条件下,塔胞藻生长和进行光合作用的最适盐度为20。高盐胁迫使塔胞藻的荧光参数Fv/Fm,Fv/Fo,ΦPSⅡ,ETR,qP均明显降低,NPQ则先升高后降低,叶绿素含量和细胞密度也显著降低。还对盐胁迫下塔胞藻的响应机制以及叶绿素荧光技术在筛选耐盐微藻品种中的应用进行了初步探讨。     

盐度胁迫下中肋骨条藻和东海原甲藻的生长与內源多胺调节作用的关系*

提要 实验室内开展了不同盐度梯度下培养常见的两种赤潮藻中肋骨条藻(Skeletonema costatum)和东海原甲藻(Prorocentrum donghaiense)的实验,根据logistic生长模型获得了生长参数Bf和mmax,并测定了藻体中过氧化物酶(POD)、二胺氧化酶(DAO)、多胺氧化酶(PAO)的活性和丙二醛(MDA)、腐胺(Put)、亚精胺(Spd)、精胺(Spm)的含量。结果表明,高、低盐度胁迫下,两种藻均会积累MDA,活性氧自由基的伤害增强,藻的生长受到了抑制。同时,两种藻都会提升多胺含量,通过多胺的调节作用来缓解胁迫的伤害,促进生长,但是在提升的多胺种类和形态上,两种藻存在着差异。高盐胁迫下,中肋骨条藻会提升多胺尤其是游离态多胺的含量来缓解伤害,东海原甲藻则依靠结合态的亚精胺和游离态的精胺的调节作用。低盐胁迫下,中肋骨条藻会提升游离态腐胺的含量,而东海原甲藻体内各形态的多胺都会上升。     

营养盐输入方式对3种微藻生长及种间竞争的影响

在不同的营养盐输入方式下(一次性输入、每2d输入、每5d输入)对杜氏盐藻(Dunaliella salina)、三角褐指藻(Phaeodactylum tricornutum)和米氏凯伦藻(Karenia mikimotoi)进行单种培养和混合培养,研究不同的营养盐输入方式对这3种海洋微藻生长及种间竞争的影响。结果表明:单种培养时,盐藻在一次性输入方式下的最终细胞密度和平均比生长速率均显著大于其它2种输入方式;三角褐指藻和米氏凯伦藻在每5d输入方式下的最终细胞密度显著大于其它2种输入方式。混合培养时,盐藻在一次性输入方式下的最终细胞密度和平均比生长速率均显著大于其它2种输入方式,最终细胞密度占总细胞密度的47.3%;三角褐指藻在一次性输入方式下的最终细胞密度和平均比生长速率均显著小于每2d输入和每5d输入方式,后2个处理组的最终细胞密度分别占总细胞密度的90.3%和79.9%;在3种输入方式下米氏凯伦藻的最终细胞密度及占总细胞密度的百分比、平均比生长速率均显著小于其它2种微藻。研究表明,无论单种培养还是混合培养,营养盐一次性输入方式均能促进盐藻的生长,而脉冲输入方式能促进三角褐指藻的生长。脉冲输入方式下混合培养时,三角褐指藻在种群中占优势。     

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.     

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。     

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.     

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.