碳酸酐酶在中肋骨条藻光合作用中的作用

探讨了在正常空气条件下生长的中肋骨条藻(Skeletonema costatum)的碳酸酐酶(CA)在其光合固碳中的作用.在中肋骨条藻的胞内和胞外均有CA活性,但胞外CA活性很低.CA抑制剂AZ(乙酰唑磺胺)对中肋骨条藻的光合放氧速率没有明显影响,而CA抑制剂EZ(乙氧苯唑胺)对其光合放氧速率有强烈的抑制作用.EZ的抑制作用使细胞最大光合速率、饱和光强和无机碳亲和力下降,无机碳的补偿点和光呼吸提高,使强光下光抑制作用增强.这些结果表明:中肋骨条藻的胞外CA在其光合作用中所起的作用较小,而其胞内CA通过催化胞内碳库中的HCO-3快速转化成CO2,提高胞内CO2的有效供给,从而提高细胞光合固碳能力和对逆境(高O2、强光和低CO2)的适应能力.     

鼠尾藻对赤潮异弯藻和中肋骨条藻的抑制作用

通过共培养的方法,研究了鼠尾藻（Surgassum thunbergii）培养水的过滤液、新鲜组织、干粉末、水溶性抽提液对赤潮异弯藻（Heterosigma akashiwo）和中肋骨条藻（Skeletonema costatum）生长的抑制效应,建立了分隔共培养系统并证明了抑制物质的存在,排除了细胞直接接触抑制的可能性.结果表明,在共培养实验中,鼠尾藻新鲜组织、干粉末及水溶性抽提液对赤潮异弯藻和中肋骨条藻的生长具有显著的抑制作用,且在较高浓度下对两种赤潮微藻的生长具有致死效应.在一次性及半连续培养方式下,鼠尾藻培养水过滤液对赤潮异弯藻的生长无抑制效应,而对中肋骨条藻的生长具有显著的抑制效应.     

赤潮水体中胶体物质对赤潮异弯藻(Heterosigma akashiwo)和中肋骨条藻(Skeletonema costatum)生长的影响

2004年2月9～10日在胶州湾发生的柔弱几内亚藻(Guinardia delicatula)赤潮期间分别在3个典型赤潮站位采集表层水样,利用过滤(0.7μm)和切向超滤(1000 Daltons)分离赤潮暴发水体中的胶体物质,分析了不同滤液及截留液的有机碳含量,并将赤潮异弯藻 (Heterosigma akashiwo)和中肋骨条藻(Skeletonema costatum)在<1000 Daltons的超滤液及具有不同含量胶体物质(1000 Daltons～0.7μm)的截留液中进行培养.结果发现在发生柔弱几内亚藻赤潮期间水体中的溶解有机碳浓度会增加,特别是胶体有机碳浓度会成倍增长;在不同胶体含量海水中培养的赤潮异弯藻的最大生长率μmax和最大生物量Bf与对照组相比会随着胶体含量的增加而显著增大,而中肋骨条藻的最大生长率μmax和最大生物量Bf与对照组相比会随着胶体含量增加而显著降低,表明柔弱几内亚藻赤潮水体中的胶体物质对赤潮异弯藻的生长起促进作用,而对中肋骨条藻的生长起抑制作用.这说明柔弱几内亚藻在增殖过程中所产生的胶体物质具有他感作用,会影响其它微藻的生长,从而可能对柔弱几内亚藻赤潮的形成起重要作用.     

尿素对中肋骨条藻与米氏凯伦藻生长的影响

采用实验室一次性培养,研究了尿素对我国东海赤潮优势藻中肋骨条藻(Skeletonema costatum)和米氏凯伦藻(Karenia mikimotoi)生长的影响。结果表明,中肋骨条藻和米氏凯伦藻均能在不同比例尿素的条件下较好地生长。随着培养液中尿素比例的增大,中肋骨条藻细胞生长速率(0.91—0.82/d)逐渐减小,平台期最大生物量(2.0×10~5—1.2×10~5个/m L)也逐渐减小,而米氏凯伦藻细胞的生长速率(0.36—0.51/d)逐渐增大,最大生物量基本不变(约1.1×10~4个/m L)。在平台期中肋骨条藻培养液中氮盐浓度最低下降到2.5μmol/L左右维持不变,而米氏凯伦藻氮盐浓度最低下降到1.0μmol/L左右。在指数生长期,随着细胞的生长溶解有机氮(DON,Dissolved Organic Nitrogen)含量迅速增加,中肋骨条藻介质中DON的浓度达到最大值(5—6μmol/L),然后浓度基本不变。米氏凯伦藻介质中DON在指数生长阶段达到最大值(2—3μmol/L)后开始下降。中肋骨条藻单细胞颗粒氮的含量(约为10~(-6)μmol,平台期约为10~(-7)μmol)要远远小于米氏凯伦藻(指数期约为10~(-4)μmol,平台期约为10~(-6)μmol)。研究表明,两种藻对尿素的吸收利用存在明显差异,在较低的溶解无机氮和较高的溶解有机氮环境中,甲藻有更好的适应性,该研究对于解释我国长江口春季硅藻和甲藻赤潮的演替有借鉴的意义。     

CO2浓度变化对拟柱胞藻生长与光合作用的影响

为了探讨大气CO₂浓度升高对水华藻类的影响,利用水华蓝藻-拟柱胞藻作为实验材料,研究了CO₂浓度升高对其生长生理和光合作用的影响,结果表明CO₂浓度升高,导致拟柱胞藻的生物量、最大光合放氧速率、光合效率显著增加。当CO₂浓度为700 mg/L以下,暗呼吸速率和光饱和点无明显影响,而CO₂浓度为1000 mg/L时,暗呼吸速率和光饱和点显著提高。随着CO₂浓度增加,藻细胞光合作用对无机碳的亲和力降低,同时胞外碳酸酐酶活性显著下降。这表明大气CO₂浓度的增加,有利于拟柱胞藻的生长和光合,进而增加了水华发生的风险。     

厦门海域2011年中肋骨条藻和血红哈卡藻赤潮期间细菌群落结构变化

【目的】研究2011年8月厦门海域爆发的由中肋骨条藻和血红哈卡藻共同引发的赤潮生消过程中细菌群落结构变化。【方法】应用变性梯度凝胶电泳技术(Denaturing gradient gel electrophoresis,DGGE)对两个赤潮站位和一个非赤潮站位的细菌群落结构进行研究。通过DGGE图谱分析确定赤潮生消过程中细菌群落中的关键菌群,借助Canoco软件分析细菌菌群与环境因子的相关性。【结果】在赤潮起始阶段细菌的群落结构与pH、N/P的相关性较大,在赤潮消亡阶段细菌的群落结构与盐度、温度呈明显的正相关。γ变形杆菌(Gammaproteobacteria)(47.7%)在赤潮期间处于主导位置,假交替单胞菌属(Pseudoalteromonas)、假单胞菌属(Pseudomonas)、交替单胞菌属(Alteromonas)、噬氢菌属(Hydrogenophaga)、Actibacter、Oleibacter等属均为优势菌群。香农-威列多样性指数表明,赤潮站位细菌多样性随着赤潮生消呈先升高后降低趋势,而非赤潮站位细菌多样性基本保持不变。通过Canoco对细菌菌群的主成分分析发现,在赤潮开始阶段Hydrogenophaga属为优势菌群,而在赤潮消退阶段则以Pseudomonas和Pseudoalteromonas属为主。赤潮站位藻际细菌和游离细菌群落多样性在藻密度较大时均达到最大值,然而两者与环境因子的相关性有较大差别。【结论】研究结果表明赤潮站位细菌群落多样性远高于非赤潮站位,细菌丰度随着赤潮藻密度的升高而增加,细菌与赤潮藻有着密切的关系。本文首次研究了多种优势藻引发的赤潮环境下细菌群落结构的变化,这对于多种藻引发的赤潮生消过程中细菌菌群结构有了深入的了解,为赤潮调控的研究提供了理论依据。     

丝藻光合作用无机碳利用研究

丝藻pH补偿点为9．2,表明其具有较低的CO2补偿点,不同pH值下丝藻光合作用放氧速率明显大于理论上从HCO3－脱水来的CO2供应速率,表明其具有利用HCO3－的能力,未检测到丝藻有胞外碳酸酐酶活性,胞内碳酸酐酶的活性随培养液中CO2浓度的升高而降低。     

粤东柘林湾中肋骨条藻(Skeletonema costatum)种群生态学

于2000年5月～2004年12月对粤东大规模养殖区柘林湾的赤潮藻中肋骨条藻（Skeletonema costatum）种群的时空分布进行了长达5a的调查。结果表明,中肋骨条藻种群密度的周年变动模式基本为双峰型,平面分布没有显著的空间差异。调查期间,中肋骨条藻种群密度的站位实测值为0～1．4×10^7cells／dm^3,总均值为3．3×10^5cells／dm^3,占浮游植物总细胞数的67．1％,为调查海区第1优势种。在总共1045份样品中,有中肋骨条藻出现的样品数为1020份。其中,种群密度大于10^6cells／dm^3的样品有65份,大于10^7cells／dm。则有4份。以大于10^6cells／dm^3为中肋骨条藻的赤潮密度标准,在调查期间至少于2000年、2003年发生4次赤潮。运用灰关联理论对中肋骨条藻种群密度与13个环境因子的关系进行排序分析发现,水温、pH值和浮游动物是影响柘林湾中肋骨条藻种群时空分布的关键因子。水温还与中肋骨条藻种群密度的对数值具极显著意义的线性关系,而达到赤潮密度的样品均落在24．5～32．0℃区间,即每年的5～9月份高温季节。由于柘林湾浮游动物的年高峰期也出现在高温季节,说明浮游动物摄食压力的存在可能是柘林湾中肋骨条藻赤潮发生的重要抑制因子。2004年调查海区中肋骨条藻种群密度和在浮游植物群落中的优势度骤然降低,可能与水体营养盐结构和Fe含量的变化有关。因此,长期调查与监测对于研究海湾生态学和赤潮发生机制是极为重要的。     

敌百虫对中肋骨条藻生长的影响

研究了有机磷农药敌百虫对中肋骨条藻Skeletonema costatum生长的影响。相对于水生甲壳类和鱼类,敌百虫对中肋骨条藻毒性较低,72 h半抑制浓度(EC₅₀)为84.8 mg/L。在31 d的亚慢性暴露下,低浓度组(≤10 mg/L)藻细胞生长趋势与对照组相似;50 mg/L的敌百虫在暴露初期对骨条藻的生长具有明显抑制作用,但在暴露15 d后,该浓度组藻细胞密度仍保持增长,最大细胞密度为2.77×10⁷ cells/mL,是对照组的137.7%。100 mg/L则能完全抑制藻细胞的生长。本实验结果说明敌百虫等有机磷农药的广泛使用可能会刺激浮游植物的生长,从而增加引发赤潮的风险。     

一株中肋骨条藻特异溶藻菌的分离鉴定及溶藻特性

【背景】赤潮频发引起严重的海洋生态学问题,不仅直接影响到海洋生态系统稳定、海洋生物资源可持续利用和水产养殖业等海洋产业的健康发展,而且对人类健康也构成了严重威胁。高效的溶藻细菌是生物法防控赤潮的有效工具之一。【目的】分离得到对中肋骨条藻具有高效溶藻效果的溶藻细菌,并对其进行分子鉴定,研究该菌株的溶藻机理以及溶藻菌所分泌溶藻物质的特性。【方法】采用2216E平板稀释涂布法分离纯化细菌,测定16SrRNA基因序列以鉴定细菌种类,利用显微镜计数溶藻菌处理后的目标藻种计算溶藻率,通过扫描电镜观察溶藻菌对中肋骨条藻的溶藻过程,利用常规生理生化方法研究溶藻菌溶藻物质的特征,并通过透析袋截留法研究溶藻物质分子量大小。【结果】分离得到一株中肋骨条藻高效溶藻菌FDHY-CJ,该菌株属于交替单胞菌属(Alteromonas sp. FDHY-CJ)。该菌株72 h处理赤潮藻结果显示,对中肋骨条藻溶藻率为95.45%,对于其他常见赤潮藻溶藻率低于40%。溶藻菌FDHY-CJ通过胞外分泌物溶藻;溶藻物质的溶藻特性不受反复冻融的影响,但对酸碱性及温度较为敏感;扫描电镜观察结果显示该溶藻菌的溶藻物质直接溶解中肋骨...     

RESPONSES TO SOLAR UV RADIATION OF THE DIATOM SKELETONEMA COSTATUM (BACILLARIOPHYCEAE) GROWN AT DIFFERENT Zn2+ CONCENTRATIONS1

Zn availability in the ocean has been suggested to limit primary production by affecting CO₂ acquisition processes for photosynthesis, therefore influencing the global carbon cycle. Also, UV radiation (UVR, 280–400 nm) is known to affect primary production in different ways. It remains to be ascertained whether Zn availability and UVR can act synergistically, antagonistically, or independently on oceanic primary production. We cultured the cosmopolitan diatom Skeletonema costatum (Grev.) Cleve under different radiation treatments with or without UVR (only photosynthetically active radiation), at 0, 3, and 10 pmol · L^?1 Zn²⁺. Specific growth rate, photosynthetic carbon assimilation, external carbonic anhydrase (eCA) activity, and estimated cell abundance increased with increasing concentrations of Zn²⁺ from 0 to 3 and 10 pmol · L^?1, irrespective of the radiation treatment. Higher eCA activity was observed in the cells grown at the high level of Zn²⁺ in the presence of UVR. An approximately linear relationship between μ and the daily dose of PAR was observed at 3 and 10 pmol · L^?1 Zn²⁺ concentrations. However, the dependency of μ on the daily PAR dose disappeared when the cells were grown in the presence of UVR, which overall depressed both μ and photosynthetic carbon assimilation. The inhibitory effect of UVR was inversely related to Zn²⁺ concentrations. The ultraviolet‐B (UVB)‐related inhibition of growth and photosynthesis decreased with time, reflecting a faster acclimation of the cells to UVR at replete Zn²⁺ levels. Overall, growth in the presence of higher Zn²⁺ concentrations reduced the sensitivity to UV radiation in Skeletonema costatum.     

COMPARISON OF CHLOROPLAST DNA AND ALLOZYME VARIATION IN WINTER STRAINS OF THE MARINE DIATOM SKELETONEMA COSTATUM(BACILLARIOPHYTA)1

Restriction fragment length polymorphisms (RFLPs) were examined in 12 winter strains of the marine diatom Skeletonema costatum (Grev.) Cleve using homologous chloroplast gene probes. The winter strains included eight different allozyme genotypes exhibiting physiological differences. These 12 winter strains were representative of the least diverse genetic group present in Narragansett Bay populations. Five chloroplast DNA probes and four different restriction enzymes were used to analyze the 12 Narragansett Bay strains and a reference strain “Skel.” A total of 46 restriction fragments were identified. All 12 of the winter strains had identical patterns. Strain Skel exhibited two RFLPs in comparison to the Narragansett Bay strains. Calculated diversity within the winter strain group was 0.0 and 0.85 for the chloroplast DNA and allozyme data, respectively. The chloroplast DNA polymorphisms revealed by this study are expected to represent a minimum level of the chloroplast DNA diversity present in Narragansett Bay seasonal populations.     

COLINEARITY OF CHLOROPLAST GENOMES IN DIVERGENT ECOTYYES OF THE MARINE DIATOM SKELETONEMA COSTATUM (BACILLARIOPHYTA)1

The chloroplast genomes of three isolates of the marine diatom Skeletonema costatum (Grev.) Cleve were mapped and found to be 131 ± 2 kb with inverted repeats (IRs) of approximately 20 kb. In contrast to higher plants, the psbA gene mapped to the IR, and rbcS mapped to the same fragment as the rbcL gene in the large single-copy region. The maps of the three isolates were colinear and revealed as many as 20 site mutations out of a total of 47 sites. The number of site mutations among isolates was consistent with previous data on their genetic diversity and physiology. Comparisons of gene order among our maps and those of three other diatom species showed that closely related genera retained similar gene orders but that more distantly related taxa exhibited extensive rearrangements. We conclude that simple restriction fragment analysis of chloroplast DNA is useful in comparative studies of diatom populations and species but that other analytical methods are more appropriate for phylogenetic studies at higher levels.     

CHARACTERIZATION OF A cDNA ENCODING GLUTAMINE SYNTHETASE FROM THE MARINE DIATOM SKELETONEMA COSTATUM (BACILLARIOPHYCEAE)

A cDNA-encoding glutamine synthetase (GS) was isolated from the marine diatom Skeletonema costatum (Greville) Cleve by PCR amplification. Nucleic acid and deduced amino acid sequences of the diatom GS were greater than 50% identical to GS from green algae and vascular plants, and phylogenetic analysis established the diatom GS as a member of the GSII gene family. The presence of an N-terminus signal sequence, identified on the basis of sequence similarity with other chloroplast-localized proteins from diatoms, suggests that the encoded GS isoenzyme is localized to the chloroplast. The GS mRNA was present in log-phase cells grown with either nitrate or ammonium as the sole added nitrogen source. Results from Southern blot analysis of genomic DNA suggested that the cDNA isolated in this study was either a member of a small, highly conserved gene family or that there was allelic variation within the region examined. Phylogenetic analyses further indicated that genes encoding GS from the diatom and two species of green algae diverged prior to the gene duplication, to the isoenzymes in vascular plants, supporting the hypothesis that GS isoenzymes in diatoms, green algae, and vascular plants arose through independent evolutionary events.     

SOURCE OF INORGANIC CARBON FOR PHOTOSYNTHESIS IN TWO MARINE DINOFLAGELLATES1

Inorganic carbon uptake was investigated in two marine dinoflagellates, Amphidinium carterae Hulburt and Heterocapsa oceanica Stein. Mass spectrometric and potentiometric assays indicated that both species lacked external carbonic anhydrase (CA). The presence of internal CA was demonstrated by potentiometric assay and by the inhibition of photosynthesis upon the addition of 500 μM ethoxyzolamide a membrane‐permeable inhibitor of CA. The capacity for bicarbonate transport was investigated by comparing the calculated rate of spontaneous CO₂ formation at pH 8.2 and 25°C with the rate of photosynthesis after the addition of 100 μM NaHCO₃. Both species appeared to have a very limited capacity for direct bicarbonate uptake. Monitoring of CO₂ and O₂ fluxes in both species by mass spectrometry demonstrated a rapid uptake of CO₂ on illumination, to concentrations below the CO₂ equilibrium concentration, indicating an effective selective uptake of CO₂. This dependence of photosynthesis on free CO₂ alone suggests that these species are CO₂ limited in their natural environment because the CO₂ concentration of seawater is very low.     

DIVERSITY IN THE GENUS SKELETONEMA (BACILLARIOPHYCEAE): III. PHYLOGENETIC POSITION AND MORPHOLOGICAL VARIABILITY OF SKELETONEMA COSTATUM AND SKELETONEMA GREVILLEI,WITH THE DESCRIPTION OF SKELETONEMA ARDENS SP. NOV.1

Skeletonema costatum (Grev.) Cleve emend. Zingone et Sarno and S. grevillei Sarno et Zingone were known only from the type material collected from Hong Kong waters more than a century ago. Both species have now been collected as live material, and their morphology and phylogenetic position are investigated in this study. Eight Skeletonema strains isolated from Florida, USA; Uruguay; and Brazil are attributed to S. costatum, while one strain from Oman is ascribed to S. grevillei based on morphological similarity to the type material of these species. In addition, a new Skeletonema species, S. ardens Sarno et Zingone, is described for a strain from Singapore and two from northern Australian waters. Skeletonema ardens has terminal fultoportula processes ending in a tapered, undulate protrusion and long intercalary fultoportulae with 1:1 junctions. The rimoportula of terminal valves is located at the margin of the valve face. No major morphological variations were observed within S. grevillei and S. ardens along a salinity gradient, whereas in S. costatum, the processes shortened and the valves came into close contact at low salinities, as already described for S. subsalsum (Cleve) Bethge. Consistent with their morphology, Skeletonema costatum and Skeletonema subsalsum also had similar rDNA sequences. Skeletonema grevillei and S. ardens were distinct in the large subunit (LSU) phylogeny. Skeletonema ardens exhibited consistent intraspecific genetic differences in both the LSU and small subunit (SSU) rDNA.     

SOURCES OF INORGANIC CARBON FOR PHOTOSYNTHESIS BY THREE SPECIES OF MARINE DIATOM1

The utilization of inorganic carbon by three species of marine diatom, Skeletonema costatum (Grev.) Cleve. Ditylum brightwellii (West) Grun., and Chaetoceros calcitrans Paulsen was investigated using an inorganic carbon isotopic disequilibnum technique and inorganic carbon dose-response curves. Stable carbon isotope data of the diatoms are also presented. Observed rates of photosynthetic oxygen evolution were greater than could be accounted for by the theoretical rate of CO₂ supply from the uncatalyzed dehydration of HCO₃^? in the external medium, suggesting use of HCO₃^? as an inorganic carbon source. Data from the isotopic disequilibrium experiment demonstrate the use of both HCO₃^? and CO₂ for photosynthesis. Carbon isotope discrimination values support the use of HCO₃^? by the diatoms.     

COMPARATIVE ALKALINE PHOSPHATASE CHARACTERISTICS OF THE ALGAL BLOOM DINOFLAGELLATES PROROCENTRUM DONGHAIENSE AND ALEXANDRIUM CATENELLA,AND THE DIATOM SKELETONEMA COSTATUM1

The alkaline phosphatase (AP) characteristics of three algal bloom species in the coastal waters of China [Prorocentrum donghaiense D. Lu, Alexandrium catenella (Whedon et Kof.) Balech, and Skeletonema costatum (Grev.) Cleve] were analyzed in a laboratory batch culture experiment using bulk assay and the single‐cell enzyme‐labeled fluorescence (ELF) method. Results showed that the AP of these three test species shared some common characteristics: AP was inducible in all three species and was expressed by algae under phosphorus (P)–stress conditions; no constitutive AP enzyme was detected in the three test species. Once AP was produced, all three test species gradually released the enzymes into the water, and the algae would reinduce AP production. There were also different specific AP characteristics among the three test species under severe P‐stressed conditions. In P. donghaiense, AP covered most of the cell, and the AP production sites were mainly on the cell surface, although some could be observed inside cells. AP also covered the whole cell of A. catenella, but the AP sites were mainly inside the cell with only some on the cell surface. Only one or two AP sites could be detected in S. costatum, and they were all on the cell surface.