Photosynthetic performance of regenerated protoplasts from disintegrated cells of Bryopsis hypnoides (Chlorophyta)

The photosynthetic performances of regenerated protoplasts of Bryopsis hypnoides, which were incubated in seawater for 1, 6, 12, and 24 h, were studied using chlorophyll (Chl) fluorescence and oxygen measurements. Results showed that for the regenerated protoplasts, the pigment content, the ratios of photosynthetic rate to respiration rate, the maximal photosystem II (PSII) quantum yield (F-v/F-m), and the effective PSII quantum yield (I broken vertical bar(PSII)) decreased gradually along with the regeneration progress, indicated that during 24 h of regeneration there was a remarkable reduction in PSII activity of those newly formed protoplasts. We assumed that during the cultivation progress the regenerated protoplasts had different photosynthetic vigor, with only some of them able to germinate and develop into mature thalli. The above results only reflected the photosynthetic features of the regenerated protoplasts at each time point as a whole, rather than the actual photosynthetic activity of individual aggregations. Further investigation suggested a relationship between the size of regenerated protoplasts and their viability. The results showed that the middle-sized group (diameter 20-60 mu m) retained the largest number of protoplasts for 24 h of growth. The changes in F-v/F-m and I broken vertical bar(PSII) of the four groups of differently sized protoplasts (i.e. < 20, 20-60, 60-100, and > 100 mu m) revealed that the protoplasts 20-60 mu m in diameter had the highest potential activity of the photosynthetic light energy absorption and conversion for several hours.

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。

Leaf orientation, incident sunlight, and photosynthesis in the alpine species Suassurea superba and Gentiana straminea on the Qinghai-Tibet Plateau

The extremely high level of solar radiation on the Qinghai-Tibet Plateau may induce photoinhibition and thus limit leaf carbon gain. To assess the effect of high light, we examined gas exchange and chlorophyll fluorescence for two species differing in light interception: the prostrate Saussurea superba and the erect-leaved Gentiana straminea. In controlled conditions with favorable water and temperature, neither species showed apparent photoinhibition in gas exchange measurements. In natural environment, however, their photosynthetic rate decreased remarkably at high light. Photosynthesis depression was aggravated under high leaf temperature or soil water stress. Relative stomatal limitation was much higher in S. superba than in G. straminea and it remarkably increased in the later species at midday when soil was dry. F-v/F-m as an indicator for photoinhibition was generally higher in S. superba than in the other species. F-v/F-m decreased significantly under high light at midday in both species, even when soil moisture was high. F-0 linearly elevated with the increment of leaf temperature in G. straminea, but remained almost constant in S. superba. Electron transport rate (ETR) increased with photosynthetically active photon flux density (PPFD) in S. superba, but declined when PPFD was high than about 1000 mumol m(-2) s(-1) in G. straminea. Compared to favorable environment, the estimated daily leaf carbon gain at PPFD above 800 mumol m(-2) s(-1) was reduced by 32% in S. superba and by 17% in G. straminea when soil was moist, and by 43% and 53%, respectively, when soil was dry. Our results suggest that the high radiation induces photoinhibition and significantly limits photosynthetic carbon gain, and the limitation may further increase at higher temperature and in dry soil.

EFFECT OF ELEVATED BICARBONATE CONCENTRATION ON GROWTH, CHLOROPHYLL A FLUORESCENCE AND ULTRA-STRUCTURE OF Microcystis aeruginosa (CYANOBACTERIUM)

The influence of bicarbonate (HCO3-) on Microcystis aeruginosa FACHB 905 was assessed in this study. Growth curves, chlorophyll a fluorescence and ultrastructure were measured at two HCO3- concentrations, 2.3 mM and 12.4 mM. A treatment of sodium chloride (NaCl) was also conducted alongside to establish the influence level of sodium. It was found that upon treatment with elevated HCO3- concentrations of 2.3 mM and 12.4 mM, cell densities were 13% and 27% (respectively) higher than controls. In photosynthetic performance, elevated HCO3- concentration initially stimulated Fv/Fm at the prophase of culture and then subsequently inhibited it. The inhibition of 2.3mM was higher than that of 12.4mM HCO3-. The maximum relative electron transport rate (ETRmax) exhibited inhibition at elevated HCO3- concentrations. DI0/CS was decreased at 2.3 mM and increased at 12.4mM. In the case of both treatments. ABS/CSI TR0/CS, ET0/CS, RC/CS0 and RC/CSm were decreased by elevated HCO3- concentrations, which indicated damage to photosynthetic apparati and an inactivation of a fraction of reaction centers. This point was also proven by ultrastructural photos. High HCO3--exposed cells lost the characteristic photosynthetic membrane arrangement compared with the control and high salinity treated samples. At the 2.3mM concentration of HCO3-. damage to photosynthetic apparati caused decreased photosynthetic activity. These findings suggested that elevated HCO3- concentration stimulated the growth and photosynthesis of M. aeruginosa FACHB 905 in a short time. Exposure to high HCO3- concentrations for a longer period of time will damage photosynthetic apparatus. In addition, the ultrastructure indicated that elevated HCO3--concentration lead to photosynthetic apparati damage. In our experiment, it was observed that the inhibition effect of 2.3mM HCO3- was higher than that of 12.4mM HCO3-. We hypothesized that M. aeruginosa FACHB 905 induced a protective mechanism under high concentrations of HCO3-.

Research on the ultrafast fluorescence property of thylakoid membranes of the wild-type and mutant rice

A high yielding rice variety mutant (Oryza sativa L., Zhenhui 249) with low chlorophyll b (Chl b) has been discovered in natural fields. It has a quality character controlled by a pair of recessive genes (nuclear gene). The partial loss of Chl b in content affects the efficiency of light harvest in a light harvest complex (LHC), thus producing the difference of the exciting energy transfer and the efficiency of photochemistry conversion between the mutant and wild-type rice in photosynthetic unit. The efficiency of utilizing light energy is higher in the mutant than that in the wildtype rice relatively. For further discussion of the above-mentioned difference and learning about the mechanism of the increase in the photochemical efficiency of the mutant, the pico-second resolution fluorescence spectrum measurement with delay-frame-scanning single photon counting technique is adopted. Thylakoid membranes of the mutant and the wild-type rice are excited by an Ar+ laser with a pulse width of 120 ps, repetition rate of 4 MHz and wavelength of 514 nm. Compared with the time and spectrum property of exciting fluorescence, conclusions of those ultrafast dynamic experiments are: 1) The speeds of the exciting energy transferred in photo-system I are faster than that in photo-system II in both samples. 2) The speeds of the exciting energy transfer of mutant sample are faster than those of the wildtype. This might be one of the major reasons why the efficiency of photosynthesis is higher in mutant than that in the wild-type rice.

Characteristics and comparative study of chlorophyll-protein complexes from siphonous green algae

By mild PAGE method, 11, 11, 7 and 9 chlorophyll-protein complexes were isolated from two species of siphonous green algae ( Codium fragile (Sur.) Harlot and Bryopsis corticulans Setch.), green alga (Ulothrix flacca (Dillw.) Thur.), and spinach (Spinacia oleracea Mill.), respectively. Apparent molecular weights, Chi a/b ratios, distribution of chlorophyll, absorption spectra, low temperature fluorescence spectra of these complexes were determined, and compared with one another. PS I complexes of two siphonous green algae are larger in apparent molecular weight because of the attachment of relative highly aggregated LHC I. Four isolated light-harvesting complexes of PSII are all siphonaxanthin-Chl a/b-protein complexes, and they are not monomers and oligomers like those in higher plants. Especially, the absence of 730 nn fluorescence in PS I complexes indicates a distinct structure and energy transfer pattern.

Fluorescence Characterization of Dissolved Organic Matter in the East China Sea after Diatom Red Tide Dispersion

Fluorescence excitation-emission spectroscopy (EEMS) was employed to analyze the 3-dimensional fluorescence of dissolved organic matter in the East China Sea after diatom red tide dispersion. The relationships between fluorescence peak intensity, and salinity and chlorophyll-a were discussed. The centers of protein-like fluorescence peaks dispersed at Ex(max)/Em(max) = 270-280/290-315 nm (Peak B), 220-230/290-305 nm (Peak D), 230-240/335-350 nm(Peak S)and 280/320 nm(Peak T). Two humic-like peaks appeared at 255-270/435-480 nm (Peak A) and 330-350/420-480 nm(Peak C). High tyrosine-like intensity was observed in diatom red tide dispersion area, and tryptophan-like fluorescence was also found which was lower. High FIB/FIS showed that diatom red tide produced much tyrosine-like matter during dispersion. Peaks S, A and C had positive correlation with one another, and their distributions were similar, which decreased with distance increasing away from the shore. Good negative correlations between peaks S, A and C and salinity suggested that Jiangsu-Zhejiang coastal water was the same source of then-L Correlations between fluorescence peak intensity and chlorophyll-a were not remarkable enough to clear the relationship between fluorescence and living algal matter. It was supposed that the living algal matter contributed little to the fluorescence intensity of algal dispersion seawater.

Comparative studies of AOT and NaDEHP by fluorescence technique and z-potential (ζ) measurements

The aggregation behaviors of two surfactants with the same hydrophobic tail, sodium bis(2-ethylhexyl)sulfosuccinate (AOT) and sodium bis(2-ethylhexyl)phosphate (NaDEHP), have been investigated by the fluorescence technique and z-potential (ζ) measurements. Five fine peaks of the pyrene molecule fluorescence spectroscopy appear in the surfactant solution, and the micropolarity at which pyrene locates is monitored from the intensity ratio of the first (I1) and the third peak (I3). A wide peak around 475 nm, the emission spectra of the excimer of pyrene molecules, is observed in the NaDEHP solution, while this is not found for the AOT system. The value of I1/I3 decreases in a more limited concentration range for the AOT system than for NaDEHP, indicating that small aggregates can be more easily formed by NaDEHP molecules. The z-potential results for the aggregates formed by the two surfactants show that the interaction between AOT and PVP is stronger than that between NaDEHP and PVP.

Simultaneous Laser-Induced Fluorescence And Contactless-Conductivity Detection For Microfluidic Chip

A combined detection system involving simultaneous LIF and contactless-conductometric measurements at the same place of the microfluidic chip was described. The LIF measurement was designed according to the confocal principle and a moveable contactless-conductivity detector was used in (CD)-D-4. Both measurements were mutually independent and advantageous in analyses of mixtures. Various experimental parameters affecting the response were examined and optimized. The performances were demonstrated by simultaneous detection of Rhodamine B. And the results showed that the combined detection system could be used sensitively and reliably. (C) 2008 Yong Yu. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Dual Confocal Laser-Induced Fluorescence/Moveable Contactless Conductivity Detector For Capillary Electrophoresis Microchip

A new dual simultaneous detector was developed for capillary electrophoresis microchip. Confocal laser-induced fluorescence (LIF) and moveable contactless conductivity detection (MCCD) were combined together for the first time. The two detection systems shared a common detection cell and could respond simultaneously. They were mutually independent and advantageous in analyses of mixtures containing organic and inorganic ions. The confocal LIF had high sensitivity and the MCCD could move along the separation channel and detect in different positions of the channel. The detection conditions of the dual detector were optimized. Rhodamine B was used to evaluate the performance of the dual detector. The limit of detection of the confocal LIF was < 5 nM, and that of the MCCD was 0.1 mu M. The dual detector had highly sensitivity and could offer response easily, rapidly and simultaneously. 

Dual confocal laser-induced fluorescence/moveable contactless

A new dual simultaneous detector was developed for capillary electrophoresis microchip. Confocal laser-induced fluorescence (LIF) and moveable contactless conductivity detection (MCCD) were combined together for the first time. The two detection systems shared a common detection cell and could respond simultaneously. They were mutually independent and advantageous in analyses of mixtures containing organic and inorganic ions. The confocal LIF had high sensitivity and the MCCD could move along the separation channel and detect in different positions of the channel. The detection conditions of the dual detector were optimized. Rhodamine B was used to evaluate the performance of the dual detector. The limit of detection of the confocal LIF was <5 nM, and that of the MCCD was 0.1 μM. The dual detector had highly sensitivity and could offer response easily, rapidly and simultaneously.

Studying atomic-resolution by X-ray fluorescence holography

In this work, the results of numerical simulations of X-ray fluorescence holograms and the reconstructed atomic images for Fe single crystal are given. The influences of the recording angles ranges and the polarization effect on the reconstruction of the atomic images are discussed. The process for removing twin images by multiple energy fluorescence holography and expanding the energy range of the incident X-rays to improve the resolution of the reconstructed images is presented. (c) 2005 Elsevier B.V. All rights reserved.