MEASUREMENT OF SPONTANEOUS EMISSION BRANCHING RATIOS IN MO(I) TRANSITIONS BY LASER-INDUCED SENSITIZED FLUORESCENCE

I. TNTRODUCTIONThe emission spectroscopic method is usually used to measure spontaneous emission branching ratios. As emission spectra cannot be detected in atomic beams, the laser-induced fluorescence or ion detection method is often used. When the fluorescence method is used to measure branching ratios, it is usually necessary to detect

FLUORESCENCE SPECTROSCOPIC STUDIES ON THE DYE MIXTURE SYSTEM C440/C540/Saf.-T

The fluorescence spectra and lifetimes of the ternary due mixture C440/C540/Saf. -T were studies. Experimental results indicated that there are efficient energy transfer among these three components of the dye system. Consequently this system may he expected to be a potemtial eandidate of laser dye giving out output in three different wavelength regions. From. the relation of donor fluorescence lifetime as a function of aceeptor concentration and the relation of accepter fluorescence intensity as a funotion...

TIDAL DEPENDENCE OF PHOTOINHIBITION OF PHOTOSYNTHESIS IN MARINE MACROPHYTES OF THE SOUTH CHINA SEA

PHOTOSYSTEM-II; CHLOROPHYLL FLUORESCENCE; ULVA-ROTUNDATA; ELECTRON-TRANSPORT; FIELD EXPERIMENTS; O-2 EVOLUTION; QUANTUM YIELD; TEMPERATURE; MACROALGAE; RESPONSES

Variations in morphology and PSII photosynthetic capabilities during the early development of tetraspores of Gracilaria vermiculophylla (Ohmi) Papenfuss (Gracilariales, Rhodophyta)

Background: Red algae are primitive photosynthetic eukaryotes, whose spores are ideal subjects for studies of photosynthesis and development. Although the development of red alga spores has received considerable research attention, few studies have focused on the detailed morphological and photosynthetic changes that occur during the early development of tetraspores of Gracilaria vermiculophylla (Ohmi) Papenfuss (Gracilariales, Rhodophyta). Herein, we documented these changes in this species of red algae. Results: In the tetraspores, we observed two types of division, cruciate and zonate, and both could develop into multicellular bodies (disks). During the first 84 hours, tetraspores divided several times, but the diameter of the disks changed very little; thereafter, the diameter increased significantly. Scanning electron microscopy observations and analysis of histological sections revealed that the natural shape of the disk remains tapered over time, and the erect frond grows from the central protrusion of the disk. Cultivation of tissue from excised disks demonstrated that the central protrusion of the disk is essential for initiation of the erect frond. Photosynthetic (i.e., PSII) activities were measured using chlorophyll fluorescence analysis. The results indicated that freshly released tetraspores retained limited PSII photosynthetic capabilities; when the tetraspores attached to a substrate, those capabilities increased significantly. In the disk, the PSII activity of both marginal and central cells was similar, although some degree of morphological polarity was present; the PSII photosynthetic capabilities in young germling exhibited an apico-basal gradient. Conclusions: Attachment of tetraspores to a substrate significantly enhanced their PSII photosynthetic capabilities, and triggered further development. The central protrusion of the disk is the growth point, may have transfer of nutritive material with the marginal cells. Within the young germling, the hetero-distribution of PSII photosynthetic capabilities might be due to the differences in cell functions.

Photosynthetic parameters of sexually different parts of Porphyra katadai var. hemiphylla (Bangiales, Rhodophyta) during dehydration and re-hydration

Physiological data from extreme habitat organisms during stresses are vital information for comprehending their survival. The intertidal seaweeds are exposed to a combination of environmental stresses, the most influential one being regular dehydration and re-hydration. Porphyra katadai var. hemiphylla is a unique intertidal macroalga species with two longitudinally separated, color distinct, sexually different parts. In this study, the photosynthetic performance of both PSI and PSII of the two sexually different parts of P. katadai thalli during dehydration and re-hydration was investigated. Under low-grade dehydration the variation of photosystems of male and female parts of P. katadai were similar. However, after the absolute water content reached 42%, the PSI of the female parts was nearly shut down while that of the male parts still coordinated well and worked properly with PSII. Furthermore, after re-hydration with a better conditioned PSI, the dehydrated male parts were able to restore photosynthesis within 1 h, while the female parts did not. It is concluded that in P. katadai the susceptibility of photosynthesis to dehydration depends on the accommodative ability of PSI. The relatively lower content of phycobiliprotein in male parts may be the cause for a stronger PSI after severe dehydration.

Study on the photosynthetic performances of Enteromorpha prolifera collected from the surface and bottom of the sea of Qingdao sea area

In this work, the photosynthetic performances of Enteromorpha prolifera thalli collected from the surface and bottom of the sea of Qingdao sea area were studied with chlorophyll fluorescence and oxygraph technology. The samples with the highest photosynthetic activity among their kinds, the floating thalli from the sea surface of the south of the Qingdao Olympic Sailing Center and the sedimentary thalli from the mud surface of the bottom Tuandao bay, were chosen as representatives of surface thalli and bottom thalli, respectively. The results showed that the maximal PSII quantum yield of the floating thalli was significantly lower than the normal level although their photosynthetic activities were relatively high; the photosynthetic potential of the thalli form the mud surface was extremely low. Thus, it is indicated that the floating thalli are seriously stressed by their environment and the thalli from the mud surface are already dead or are dying. On the other hand, the results of the laboratory cultivation showed that the sedimentary thalli cannot regain normal photosynthetic activity even under normal illumination conditions. Thus, the thalli from the mud surface cannot become reproductive source of the alga even if they can reach sea surface again. Therefore, a preliminary conclusion can be reached that, up to mid-July 2008, the environmental conditions of the Qingdao sea area are not suitable for the growth of the alga E. prolifera and for this reason the biomass of E. prolifera, in the area, could be declining.

Characterization of the main light-harvesting chlorophyll a/b-protein complex of green alga, Bryopsis corticulans

The main light-harvesting chlorophyll a/b -protein complex (LHC II) has been isolated directly from thylakoid membranes of shiphonous green alga, Bryopsis corticulans Setch. by using two consecutive runs of anion exchange and gel-filtration chromatography. Monomeric and trimeric subcomplexes of LHC 11 were obtained by using sucrose gradient ultracentrifugation. Pigment analysis by reversed-phase high performance liquid chromatography showed that chlorophyll a (Chl a), chlorophyll b (Chl b), neoxanthin, violaxanthin and siphonaxanthin were involved in LHC 11 from B. corticulans. The properties of electronic transition of monomeric LHC II showed similarities to those of trimeric LHC II. Circular dichroism spectroscopy showed that strong intramolecular interaction of excitonic dipoles between Chl a and between Chl b exist in one LHC II apoprotein, while the intermolecular interaction of these dipoles can be intensified in the trimeric structure. The monomer has high efficient energy transfer from Chl b and siphonaxanthin to Chl a similarly to that of the trimer. Our results suggest that in B. corticulans, LHC II monomer has high ordered pigment organization that play effective physiological function as the trimer, and thus it might be also a functional organization existing in thylakoid membrane of B. corticulans.

Isolation of the main light-harvesting chlorophyll a/b-protein complex from thylakoid membranes of marine alga, Bryopsis corticulans by a direct method

The main chlorophyll a/b light-harvesting complex (LHC 11) has been isolated directly from thylakoid membranes of marine green alga (Bryopsis corticulans Setch.) by two consecutive runs of anion exchange and gel-filtration chromatography. LHC 11 proteins in the membrane extracts treated with 3% n-Octyl-b-D-glucopyranoside (OG) obtained specific binding ability on Q Sepharose column, and thus were isolated from the thylakoid membranes in a highly selective fraction. The monomeric, trimeric and oligomeric subcomplexes of LHC 11 have been obtained by fractionation of the LHC 11 mixes with sucrose density gradient ultracentrifugation. The SDS-PAGE analysis of peptide composition and absorption spectrum showed that LHC 11 monomers, trimers and oligomers prepared through this work were intact and in high purity. Our report is the first to show that it is possible to purify LHC If directly from thylakoid membranes without extensively biochemical purification.

A centromeric satellite sequence in the Pacific oyster (Crassostrea gigas Thunberg) identified by fluorescence in situ hybridization

A highly repetitive satellite sequence was previously identified in the Pacific oyster Crassostrea gigas Thunberg. The sequence has 168 bp per unit, present in tandem repeats, and accounts for 1% to 4% of the genome. We studied the chromosomal location of this satellite sequence by fluorescence in situ hybridization (FISH), A probe was made by polymerase chain reaction and incorporation of digoxigenin-11-dUTP. Hybridization was detected with fluorescein-labeled antidigoxigenin antibodies. FISH signals were located at centromeric regions of 7 pairs of the Pacific oyster chromosomes. No interstitial site was found. Signals were strong and consistent on chromosomes 1, 2, 4, and 7, but weak or variable oil chromosomes 5, 8, and 10. No signal was observed on chromosomes 3, 6, and 9. Our results showed that this sequence is clearly a centromeric satellite, disputing its previous assignment to the telomeric and submetacentric regions of 2 chromosomes. No signal was detected in the American oyster (Crassostrea virginica Gmelin).

Development of rRNA and rDNA-targeted probes for fluorescence in situ hybridization to detect Heterosigma akashiwo (Raphidophyceae)

Heterosigma akashiwo (Hada) is a fragile, fish-killing alga. Efforts to understand and prevent blooms due to this harmful species to mitigate the impact on aquaculture require the development of methods for rapid and precise identification and quantification, so that adequate warning of a harmful algal bloom may be given. Here, we report the development and application of rRNA and rDNA-targeted oligonucleotide probes for fluorescence in situ hybridization (FISH) to aid in the detection and enumeration of H. akashiwo. The designed probes were species specific, showing no cross-reactivity with four common HAB causative species: Prorocentrum micans Ehrenberg, P. minimum (Pavillard) Schiller, Alexandrium tarmarense (Lebour) Balech, and Skeletonema costatum (Greville) Cleve, or with four other microalgae, including Gymnodinium sp. Stein, Platy-monas cordiformis (Karter) Korsch, Skeletonema sp.1 Greville and Skeletonema sp.2. The rRNA-targeted probe hybridized to cytoplasmic rRNA, showing strong green fluorescence throughout the whole cell, while cells labeled by rDNA-targeted probe exhibited exclusively fluorescent nucleus. The detection protocols were optimized and could be completed within an hour. For rRNA and rDNA probes, about a corresponding 80% and 70% of targeted cells could be identified and quantified during the whole growth circle, despite the inapparent variability in the average probe reactivity. The established FISH was proved promising for specific, rapid, precise, and quantitative detection of H. akashiwo. (C) 2008 Elsevier B.V. All rights reserved.

Isolation and mapping of telomeric pentanucleotide (TAACC)(n) repeats of the pacific whiteleg shrimp, Penaeus vannamei, using fluorescence in situ hybridization

To develop genetic and physical maps for shrimp, accurate information on the actual number of chromosomes and a large number of genetic markers is needed. Previous reports have shown two different chromosome numbers for the Pacific whiteleg shrimp, Penaeus vannamei, the most important penaeid shrimp species cultured in the Western hemisphere. Preliminary results obtained by direct sequencing of clones from a Sau3A-digested genomic library of P. vannamei ovary identified a large number of (TAACC/GGTTA)-containing SSRs. The objectives of this study were to (1) examine the frequency of (TAACC)(n) repeats in 662 P. vannamei genomic clones that were directly sequenced, and perform homology searches of these clones, (2) confirm the number of chromosomes in testis of P. vannamei, and (3) localize the TAACC repeats in P. vannamei chromosome spreads using fluorescence in situ hybridization (FISH). Results for objective I showed that 395 out of the 662 clones sequenced contained single or multiple SSRs with three or more repeat motifs, 199 of which contained variable tandem repeats of the pentanucleotide (TAACC/GGTTA),, with 3 to 14 copies per sequence. The frequency of (TAACC)n repeats in P. vannamei is 4.68 kb for SSRs with five or more repeat motifs. Sequence comparisons using the BLASTN nonredundant and expressed sequence tag (EST) databases indicated that most of the TAACC-containing clones were similar to either the core pentanucleotide repeat in PVPENTREP locus (GenBank accession no. X82619) or portions of 28S rRNA. Transposable elements (transposase for Tn1000 and reverse transcriptase family members), hypothetical or unnamed protein products, and genes of known function such as 18S and 28S rRNAs, heat shock protein 70, and thrombospondin were identified in non-TAACC-containing clones. For objective 2, the meiotic chromosome number of P. vannamei was confirmed as N = 44. For objective 3, four FISH probes (P1 to P4) containing different numbers of TAACC repeats produced positive signals on telomeres of P. vannamei chromosomes. A few chromosomes had positive signals interstitially. Probe signal strength and chromosome coverage differed in the general order of P1 > P2 > P3 > P4, which correlated with the length of TAACC repeats within the probes: 83, 66, 35, and 30 bp, respectively, suggesting that the TAACC repeats, and not the flanking sequences, produced the TAACC signals at chromosome ends and TAACC is likely the telomere sequence for P. vannamei.

Isolation and characterization of the oxygen-evolving photosystem II complex from the economical red alga Bangia fusco-purpurea

The highly pure and active photosystem II (PSII) complex was isolated from Bangia fusco-purpurea (Dillw) Lyngb., an important economic red alga in China, through two steps of sucrose density gradient ultracentrifugation and characterized by the room absorption and fluorescence emission spectra, DCIP (2,6-dichloroindophenol) reduction, and oxygen evolution rates. The PSII complex from B. fusco-purpurea had the characteristic absorption peaks of chlorophyll (Chl) a (436 and 676 nm) and typical fluorescence emission peak at 685 nm (Ex = 436 nm). Moreover, the acquired PSII complex displayed high oxygen evolution (139 mu mol O-2/(mg Chl h) in the presence of 2.5 mM 2,6-dimethybenzoqinone as an artificial acceptor and was active in photoreduction of DCIP (2,6-dichloroindophenol) by DPC (1,5-diphenylcarbazide) at 163 U/(mg Chl a h). SDS-PAGE also suggested that the purified PSII complex contained four intrinsic proteins (D1, D2, CP43, and CP47) and four extrinsic proteins (33-kD protein, 20-kD protein, cyt c-550, and 14-kD protein).

中国近海浮游植物光合作用研究——初级生产力模型计算与活体叶绿素荧光测量

本文的研究内容是初级生产力模型计算方法以及活体叶绿素荧光测量方法在我国近海浮游植物光合作用研究中的应用。 论文对在我国近海实测的1321 组叶绿素垂直分布数据以及84 组初级生产力数据进行了非线性拟合分析，得出了描述叶绿素和初级生产力垂直变化规律的相应参数。结合光合参数——初始斜率α 的平面分布将我国近海分为9 个光合作用特征相对独立的分区。以光强和遥感叶绿素为输入变量，以各分区实测参数的平均值为模型参数，首次建立了分海区的时间－深度积分初级生产力计算模型。 对拟合参数的分析表明：我国近海深水叶绿素峰（DCM）在夏季最明显而在冬季则不明显，这与夏季温跃层的形成和冬季温跃层消失的规律相符合。最大光合速率参数主要受温度和营养盐状况的影响，但整体而言各个季节的变化不大。光合作用初始斜率参数主要受浮游植物光照条件和光照历史的影响，从春季到冬季持续增加，基本与光照强度的变化刚好相反。光抑制参数反映着浮游植物对高光强伤害的敏感性，它在夏、秋季较低而在冬季较高。 模型计算结果表明我国近海初级生产力规模约为6.4±1.0×108tC/a。总体来说我国近海海域初级生产力北方高于南方，西部高于东部。北方初级生产力高峰期大都出现在夏季而南方出现在冬季，中部则高峰期出现在春秋两季，与温跃层出现和消失的规律有一定的相关性。与由实测生产力推算的初级生产力相比，本模型的计算结果较高，误差产生的原因包括二类水体遥感叶绿素浓度偏高、地形因素等。与VGPM相比较，本模型的计算结果更接近于实测结果。模型中云覆盖度参数的变化可能引起计算结果约25％的变化，当取云覆盖度参数为0.5 时的计算结果与用实际天气状况数据的计算结果较为接近。制作了基于分区模型的水柱初级生产力计算软件，可在仅测量叶绿素浓度的情况下进行水柱初级生产力的计算。 模型准确度的进一步提高的途径主要包括实测参数数量的增加以及遥感叶绿素浓度准确性的提高。 与初级生产力相比，活体叶绿素荧光测量可以更快速、简便地跟踪浮游植光合作用动态。本研究首次将活体叶绿素荧光测量方法应用于野外浮游植物光合作用状态研究。通过暗驰豫实验，建立了使用OS5－AFM藻类荧光仪现场测定活体叶绿素荧光参数——Fv/Fm（光化学效率）的操作方法，并对胶州湾海域浮游植物光化学效率进行了高时空分辨率的调查，对相关因素进行了分析，并尝试通过Fv/Fm进行初级生产力计算。 结果表明，胶州湾海域表层Fv/Fm值的全年平均值约0.37，处于中等偏低水平。Fv/Fm值在秋季最高而春季最低，其平面分布在夏季较均匀而在春季则有较大的区域差异。湾东部Fv/Fm值变化幅度较大，一年中有4 个高峰期；而湾西部Fv/Fm值变化幅度较小，一年中有3 个高峰期。光合作用最活跃的海区随季节更替而变化，其年变化模式可能具有一定的重复性。不同深度上Fv/Fm值的海区平均值没有显著差异，但底层Fv/Fm值的空间差异较小。 通过对营养盐和Fv/Fm值时间变化的分析，首次发现了自然水体中Fv/Fm值变化对氮盐浓度变化响应的滞后效应，在不同的站点，影响Fv/Fm值的氮盐种类有所不同，湾西部海域与NH4+相关性较高而湾东部海域与NO3-的相关性较高，P、Si等则与Fv/Fm值基本无相关性。滞后期的长度并不稳定，1 旬和2 旬的滞后现象都可观察到。Fv/Fm值变化率与氮盐浓度变化率之间的相关性高于Fv/Fm值与氮盐浓度之间的相关性。这表明即使在胶州湾这样无机氮浓度很高、浮游植物生长基本不受氮源限制的水体，氮源浓度的变化仍然影响着浮游植物的光合作用状态。 初步建立了由Fv/Fm测量结果计算初级生产力的方法。模型的计算结果与文献资料中胶州湾水域初级生产力的时空分布趋势基本一致，但在数量上明显低于传统估算的结果。获得更多同步测量的初级生产力和Fv/Fm值数据可逐渐改善这一方法计算结果的准确性，并可籍次方法更快更方便地进行初级生产力的测算。

海洋微藻产生溶解有机物及东海溶解有机物三维荧光特征研究

本文通过胶州湾围隔实验、微藻培养实验研究了海洋微藻产生溶解有机物的三维荧光特性，并初步利用经验正交函数对三维荧光光谱图进行了主成分分析，通过东海取样研究了东海荧光溶解有机物的时空分布特征，并对荧光溶解有机物的来源进行了分析。得到了一些初步结论： 胶州湾围隔实验中不同营养盐培养结果表明浮游植物可产生类蛋白和类腐殖质荧光，类蛋白荧光峰由类酪氨酸(tyrosine-like)荧光峰和类色氨酸(tryptophan-like)荧光峰组成，主要位置为Exmax/Emmax=270nm/290~310nm，Exmax/Emmax= 270~290/320~350的荧光峰强度比较弱；在Exmax/Emmax=250~260/380~480nm(A峰)、Exmax/Emmax=310~320/380~420nm(C峰)和Exmax/Emmax=330~350/420~480nm(M峰)位置均出现零散的类腐殖质荧光峰，其中以A峰为主。类酪氨酸荧光强度明显高于类腐殖质荧光强度。浮游植物量降低时，类酪氨酸荧光强度与叶绿素a浓度呈明显的负相关。硅藻和甲藻产生的类酪氨酸和类色氨酸荧光强度之间具有较好的相关性，两者来源相似, 并且甲藻与硅藻相比能够产生更多的类蛋白荧光物质。不同环境下类腐殖质混合物的组分比例不同，甲藻生长环境下相对于硅藻具有较低的A:C比值。 在实验室培养中肋骨条藻（Skeletonema costatum）、塔玛亚历山大藻（Alexandrium tamarense）、微小亚历山大藻（Alexandrium mimutum）、锥状斯氏藻（Scrippsiella trochoidea）、东海原甲藻（Prorocentrum donghaiense）及海洋原甲藻（Prorocentrum micans）的实验结果表明，微藻在生长过程中产生出荧光溶解有机物，中肋骨条藻为代表的硅藻主要产生类腐殖质荧光物质，而甲藻在指数增长期主要产生类蛋白荧光物质。进入消亡期后类蛋白荧光和类腐殖质荧光强度迅速增大，原因可能是衰老、死亡藻细胞的破碎释放出大量的荧光有机物质所致，此外还有细菌对非荧光有机物进一步降解。塔玛亚历山大藻、微小亚历山大藻、东海原甲藻及海洋原甲藻的类蛋白荧光强度在消亡后期由于细菌降解或光降解等因素而降低。同属微藻产生的荧光物质相似，例如塔玛亚历山大藻与微小亚历山藻、东海原甲藻与海洋原甲藻，但具体荧光峰位置有所不同。利用经验正交函数能够对三维荧光光谱谱图进行主成份分析。 在利用三维荧光光谱法研究长江口海域台风前后不同站位荧光溶解有机物荧光特性及分布特征的结果表明，长江口海区主要的荧光溶解有机物荧光峰为T峰、S峰和A峰。风前和风后的类色氨酸分别来源于相似物质。风前，在表层浮游植物能够产生类蛋白荧光物质，而底层类蛋白荧光物质不受浮游植物的影响，长江冲淡水能够带来部分类蛋白荧光物质；表层的类腐殖质不受浮游植物的影响，而底层的浮游植物在降解过程中能够产生一部分类腐殖质，并且长江冲淡水对表层和底层的类腐殖质来源均有很大贡献。风后，表层的类色氨酸与叶绿素a浓度不呈相关性，而底层却呈正相关，另外表层和底层的类色氨酸均受到长江冲淡水的影响；表层和底层的类腐殖质与叶绿素a均没有相关性，但受陆源影响显著，长江冲淡水能带来类腐殖质。

A field study on the conversion ratio of phytoplankton biomass carbon to chlorophyll-a in Jiaozhou Bay, China

A one-year field study was conducted to determine the conversion ratio of phytoplankton biomass carbon (Phyto-C) to chlorophyll-a (Chl-a) in Jiaozhou Bay, China. We measured suspended particulate organic carbon (POC) and phytoplankton Chl-a samples collected in surface water monthly from March 2005 to February 2006. The temporal and spatial variations of Chl-a and POC concentrations were observed in the bay. Based on the field measurements, a linear regression model II was used to generate the conversion ratio of Phyto-C to Chl-a. In most cases, a good linear correlation was found between the observed POC and Chl-a concentrations, and the calculated conversion ratios ranged from 26 to 250 with a mean value of 56 A mu g A mu g(-1). The conversion ratio in the fall was higher than that in the winter and spring months, and had the lowest values in the summer. The ratios also exhibited spatial variations, generally with low values in the near shore regions and relatively high values in offshore waters. Our study suggests that temperature was likely to be the main factor influencing the observed seasonal variations of conversion ratios while nutrient supply and light penetration played important roles in controlling the spatial variations.