CHLOROPHYLL FLUORESCENCE RATIO F685/F735 IN BROWN ALGAE AND ITS VARIATION UNDER EXCITATION BY TWO TYPES OF LIGHT AND DEHYDRATION

Calculation and comparative study of the chlorophyll fluorescence ratio F685/F735 in brown algae (Laminaria japonica, Underia pinnatifida and Padina crassa) excited by blue and green light showed that the fluorescence ratios were higher when the algae were excited by blue light (440 nm), but reduced obviously under green light (540 nm) excitation. The values also reduced under dehydration but could recover during rehydration if the stress was not serious. The variation of the fluorescence ratio under dehydration was mainly because changes in fluorescence emission at 735 nm were always sharper than those at 685 nm. The ratio was sensitive to stress and has potential as a stress indicator in phycological research. Measurement of the fluorescence excitation spectra showed that the only peak at 540 hm changed apparently during dehydration. It meant that the function of the Chl a /Fucoxanthin protein complex for energy transfer was easily inhibited by water stress. However, no variation of the ratio was foun     

Comparative studies on spectral properties of R-phycoerythrin from the red seaweeds from Qingdao

The present paper reports on comparative studies of R-phycoerythrin (R-PE) from 30 species of the red seaweeds in Bangiophyceae and in Florideophyceae from Qingdao. The measured absorption spectra show that R-PE from these red seaweeds have two spectral types, namely type I having two absorption peaks and type II having three absorption peaks. 9 specise of a lower red seaweed belonging to Bangiophyceae are all type I R-PE. 3 species of a higher red seaweed belonging to Florideophyceae are also type I R-PE. The remaining 18 species are all type II R-PE. According to the distribution of two types of R-PE in red algae, an evolutionary trend from type I R-PE to type II R-PE, can be seen, but this evolutionary process is more complicated and without a definite line of demarcation between the two spectral types of R-PE. The occurrence of two types of R-PE in red algae is of taxonomic significance. Project supported by the Science Fund of the Chinese Academy of Sciences. Contribution No. 1357 from the Institute of Oceanology, Academia Sinica.     

EFFECTS OF DCMU ON CHLOROPHYLL FLUORESCENCE RATIO F685/F735 IN MARINE RED,BROWN AND GREEN ALGAE

The chorophyll fluorescence ratio F685/F735 in vivo can be a useful indicator for stress detection inhigher plants and seaweed. DCMU [3 - (3, 4 - dichlorophenyl ) - 1, 1 - dimethylurea] treatment influencesthis ratio. Th effects of DCMU on F685/F735 of marine red, brown and green algae under excitationlight of different wavelengths were investigted. In the brown algae, Laminaria japonica and Undariapinnatifida,DCMU did not increase this ratio under blue light excitation but increased the ratio slightlyunder excitation by green light. For the red algea, Halymenia sinensis, DCMU increased the ratiomarkedly under both blue and green ligh excitation. The percentage increase could reach 50% (undergreen light excitation) and was due to unequal enhancement at the two emission maxima by DCMU. Afraction of chlorophyll which contributed to fluorescence in the 735 nm region was less sensitive toDCMU and was likely from photosystem I of red algae. In the green alga, Ulva pertusa, DCMU causeda slight increase in F685     

Fluorometric discrimination technique of phytoplankton population based on wavelet analysis

The discrete excitation-emission-matrix fluorescence spectra (EEMS) at 12 excitation wavelengths (400, 430, 450, 460, 470, 490, 500, 510, 525, 550, 570, and 590 nm) and emission wavelengths ranging from 600-750 nm were determined for 43 phytoplankton species. A two-rank fluorescence spectra database was established by wavelet analysis and a fluorometric discrimination technique for determining phytoplankton population was developed. For laboratory simulatively mixed samples, the samples mixed from 43 algal species (the algae of one division accounted for 25%, 50%, 75%, 85%, and 100% of the gross biomass, respectively), the average discrimination rates at the level of division were 65.0%, 87.5%, 98.6%, 99.0%, and 99.1%, with average relative contents of 18.9%, 44.5%, 68.9%, 73.4%, and 82.9%, respectively; the samples mixed from 32 red tide algal species (the dominant species accounted for 60%, 70%, 80%, 90%, and 100% of the gross biomass, respectively), the average correct discrimination rates of the dominant species at the level of genus were 63.3%, 74.2%, 78.8%, 83.4%, and 79.4%, respectively. For the 81 laboratory mixed samples with the dominant species accounting for 75% of the gross biomass (chlorophyll), the discrimination rates of the dominant species were 95.1% and 72.8% at the level of division and genus, respectively. For the 12 samples collected from the mesocosm experiment in Maidao Bay of Qingdao in August 2007, the dominant species of the 11 samples were recognized at the division level and the dominant species of four of the five samples in which the dominant species accounted for more than 80% of the gross biomass were discriminated at the genus level; for the 12 samples obtained from Jiaozhou Bay in August 2007, the dominant species of all the 12 samples were recognized at the division level. The technique can be directly applied to fluorescence spectrophotometers and to the developing of an in situ algae fluorescence auto-analyzer for phytoplankton population.     

Measurement and feature analysis of absorption spectra of four algal species

Two methods for particulate pigments(i.e.,quantitative filter technique,QFT,and in vivo measurement,InVivo,respectively)and two methods for dissolved pigments(i.e.,Acetone Extracts,AceEx,and high-performance liquid chromatography,HPLC,respectively) were used to obtain the optical absorption coefficient spectra for cultures of four typical algal species.Through normalization and analysis of the spectra,it is shown that(1) the four methods are able to measure optical absorption spectra of particulate and/or dissolved pigments;(2)that the optical absorption spectra of particulate and dissolved pigments were consistent in terms of the peak position in the blue wavelength,and the difference of the peak position in the near infrared wavelength was ~10nm between each other;and(3)that the leveling effect of the absorption spectra of particulate pigments was significant.These four methods can all effectively measure the absorption coefficients of phytoplankton pigments,while each one has its unique advantages in different applications.Therefore,appropriate method should be carefully selected for various application due to their intrinsic difference.     

Study on Optical Properties of Unpigmented Suspended Particles,Yellow Substance and Phytoplankton Algae in Taihu Lake

The optical properties of a waterbody are subjects of research on limnological hydrooptics. The optical properties of, and parameterization methods for, three impurities in Taihu Lake, namely unpigmented suspensions, yellow substance and phytoplankton algae, are analyzed in detail in this paper dealing with the optical types of Taihu Lake waters both in winter and in summer. The results showed that : 1 ) The optical parameters of suspended particles can be calculated by the application of Mie Theory; 2) The absorption of yellow substance decreases exponentially with increasing wavelength, with the mean value of decreasing rate being about 0.014 nm^-1 ; 3) Taihu Lake waters appeared to be of type PY in winter, with suspended particles and yellow substance being the dominant light absorbers, and type CPY in summer, with suspended particles, yellow substance and phytoplankton algae being the dominant light absorbers; 4) In winter or summer, unpigmented suspension has the main effect on the scattering property of Taihu Lake waters.     

ISOLATION, PURIFICATION AND CHARACTERIZATION OF THE HYDROGEN EVOLUTION PROMOTING FACTOR OF HYDROGENASE OF SPIRULINA PLATENSIS

A component (s-factor) with obvious promoting effect on hydrogen evolution of hydrogenase has beenisolated and extracted from a Cell=free preparation of Spirulina platensis.The effect of the s-factor in the re-action system is similar to that of Na_2S_2O_4, but is coupled with ligh. The s-factor has the maximumabsorption peak at 620 nm in the oxidized state, at 590 nm in the reduced state. The partially purifieds-factor showed two bands by SDS-PAGE and is distinctly different from phycocyanin,which has nochange of oxidized state and reduced state absorption spectra, and also has no promoting effect onhydrogenase of Spirulina platensis under the light.     

The isolation of prophyra-334 from marine algae and its UV-absorption behavior

1 INTRODUCTIONSolar ultraviolet radiation on the earth's surfaceincreases with the depletion of stratospheric ozone(Crutzen, 1992; Kerr and McElroy, 1993; Mad-ronich et al., 1998). Ultraviolet radiation is a highlyactive component of solar spectrum. The excessiveUV radiation has deleterious effects on all plant,animal and microbial groups exposed to solar radia-tion, including the destruction of DNA, proteins andother molecules (Harm, 1980), inhibition of photo-synthesis and growth (W…     

Isolation,purification and characterization of the hydrogen evolution promoting factor of hydrogenase ofSpirulina platensis

A component (s-factor) with obvious promoting effect on hydrogen evolution of hydrogenase has been isolated and extracted from a cell-free preparation ofSpirulina platensis. The effect of the s-factor in the reaction system is similar to that of Na₂S₂O₄, but is coupled with light. The s-factor has the maximum absorption peak at 620 nm in the oxidized state, at 590 nm in the reduced state. The partially purified s-factor showed two bands by SDS-PAGE and is distinctly different from phycocyanin, which has no change of oxidized state and reduced state absorption spectra, and also has no promoting effect on hydrogenase ofSpirulina platensis under the light. The research supported by the NSFC.     

Multiple excitation wavelength fluorescence emission spectra technique for discrimination of phytoplankton

In vivo fluorescence methods are efficient tools for studying the distribution of phytoplankton in nature.Different algae species usually have different pigments with different ratios,which results in different fluorescence emission spectra.Based on multiple excitation wavelength fluorescence emission spectra,a discrimination technique is established in this study.The discrimination method,established by multivariate linear regression and weighted least-squares,was used to differentiate the samples cultured in the laboratory and collected from Jiaozhou Bay near Qingdao at the division level.The correctly discriminated samples were ≥ 86% for single algae samples,≥ 88% for simulatively mixed ones,≥ 91% for physically mixed ones and 100% for samples collected from Jiaozhou Bay.The result in this research is more definite for the physically mixed samples in the laboratory.The method described here can be employed to monitor the phytoplankton population in the marine environment.     

引进印度尼西亚大珠母贝人工育苗试验

在雷州半岛海域水温24.0～30.0℃、盐度30.8～32.5、pH8.0～8.3条件下,以牟氏角毛藻,亚心形扁藻,牟氏角毛藻、亚心形扁藻和小球藻,天然混合藻等4种饵料,进行印度尼西亚产大珠母贝亲贝促熟培育试验。结果表明:不同饵料组合在亲贝促熟百分率、排精产卵率和受精率之间差异不显著(P>0.05);阴干温差流水刺激可以有效诱导亲贝排精产卵,优选幼虫和投放合适的采苗器可以提高幼虫的变态率。     

Researches on the stability of porphyra-334 solution and its influence factors

1　Introduction　Ultravioletradiationisahighlyactivecomponentofsolarspectrum .Ultravioletradiationhasdetrimentaleffectsonlivingthingsexposedtosunlight ,includingthedestructionofDNA ,proteinsandothermolecules(Harm ,1980 ) ,inhibitionofphotosynthesisandgrow…     

Chlorophyll fluorescence extraction from water-leaving radiance of algae-containing water through polarization

When measuring reflectance spectra, it is very important to accurately extract chlorophyll fluorescence from elastic- scattering light in water-leaving radiance. The elastic scattering of light by water particles produces partially polarized light. In contrast, chlorophyll fluorescence in planktonic algae yields completely unpolarized light. These properties can be used to separate fluorescent signals from the water-leaving radiance and thus to determine chlorophyll concentration. The algal species Aureococcus anophagefferens was used to conduct a laboratory polarization experiment. For the tests, we used a field spectroradiometer and a polarizer; measurements were collected using two different observation modes. The chlorophyll fluorescence curve extracted through polarization shows an excellent match with the results obtained using the fluorospectro photometer for both measurement modes, suggesting that polarization-based chlorophyll fluorescence extraction may be feasible. The extracted fluorescence is more reliable at incident zenith angles ranging from 30° to 60°. For algae-containing water, the results improve with increasing chlorophyll concentration. This method could help improve chlorophyll concentration measurement and the remote-sensing detection of resulting harmful algae blooms.     

分频输电系统中变频变压器的两种新型结构

分析了星-开口三角型三相铁磁三倍叛变压器的移相原理和用于分频输电系统时可能出现的问题，提出了两种新型的铁磁三倍频变压器的结构方案，并论述了它们的工作原理和优点。指出对铁磁三倍频变压器仍需进行更深入研究的若干问题．     

Three-dimensional fluorescence characteristics of dissolved organic matter produced by <Emphasis Type="Italic">Prorocentrum donghaiense</Emphasis> Lu

Filtration and cross-flow ultrafiltration techniques were used to separate culture media of Prorocentrum donghaiense at the exponential growth,stationary and decline stages into <0.45 μm filtrate,100 kDa-0.45 μm,10-100 kDa and 1-10 kDa retentate and <1 kDa ultrafiltrate fractions.The fluorescence properties of different molecular weights of dissolved organic matter(DOM) were measured by excitation-emission matrix spectra.Protein-like and humic-like fluorophores were observed in the DOM produced by P.donghai...     

Accurate quantification of astaxanthin from Haematococcus crude extract spectrophotometrically

The influence of alkali on astaxanthin and the optimal working wave length for measurement of astaxanthin from Haematococcus crude extract were investigated, and a spectrophotometric method for precise quantification of the astaxanthin based on the method of Boussiba et al. was established. According to Boussiba's method, alkali treatment destroys chlorophyll. However, we found that: 1) carotenoid content declined for about 25% in Haematococcus fresh cysts and up to 30% in dry powder of Haematococcus broken cysts after alkali treatment; and 2) dimethyl sulfoxide (DMSO)-extracted chlorophyll of green Haematococcus bares little absorption at 520-550 nm. Interestingly, a good linear relationship existed between absorbance at 530 nm and astaxanthin content, while an unknown interference at 540-550 nm was detected in our study. Therefore, with 530 nm as working wavelength, the alkali treatment to destroy chlorophyll was not necessary and the influence of chlorophyll, other carotenoids, and the unknown interference could be avoided. The astaxanthin contents of two samples were measured at 492 nm and 530 nm; the measured values at 530 nm were 2.617 g/100 g and 1.811 g/100 g. When compared with the measured values at 492 nm, the measured values at 530 nm decreased by 6.93% and 11.96%, respectively. The measured values at 530 nm are closer to the true astaxanthin contents in the samples. The data show that 530 nm is the most suitable wave length for spectrophotometric determination to the astaxanthin in Haematococcus crude extract.     

海南省三亚海藻资源调查

通过对海南島三亚海藻的分布和种类进行深入调查,采集和整理了标本123号,计35属53种,并分析了三亚的气候和环境条件对海藻的生长与分布的关系,为三亚海藻的开发与利用提供了第一手资料。     

Subunit composition and chromophore content of R-phycoerythrin fromPorphyra haitanensis

R-phycoerythrin fromPorphyra haitanensis exists in two aggregation states with different molecular weights. A more highly aggregated form, RPE I, was chromatographed on Bio-Rex 70 column with urea solution (pH 3.0) as eluent, and the molecular weights of the 3 subunits (α, β, γ) obtained were determined on SDS-PAGE at 18000, 19200 and 30000, respectively. α subunit carried two phycoerythrobilin (PEB); β subunit, three PEB and one phycourobilin (PUB); γ subunit, one PEB and three PUB chromophores. The molar ratio of α, β, and γ subunits of RPE I was 6: 6: 1, and their subunit composition was confired to be (αβ)₆γ on account of the molecular weight of RPE I, 232000. A lower aggregated form, RPE II, contained α and β subunits similar to those of RPE I, but its subunit composition was the (αβ) monomer of RPE. Contribution No. 2108 from the Institute of Oceanology, Chinese Academy of Sciences     

Cloning and characterization of glyceraldehyde-3-phosphate dehydrogenase encoding gene inGracilaria/Gracilariopsis lemaneiformis

Glyceraldehyde-3-phosphate dehydrogenase （GAPDH） plays important roles in various cellular processes. A cytosolic GAPDH encoding gene （gpd） of Gracilaria/Gracilariopsis lemaneiformis was cloned and characterized. Deduced amino acid sequence of the enzyme of G. lemaneiformis had high homology with those of seven red algae. The 5＇-untranslated regions of the GAPDHs encoding genes of these red algae varied greatly. GAPDHs of these red algae shared the highly conserved glyceraldehyde 3-phosphate dehydrogenase active site ASCTTNCL. However, such active site of Cyanidium caldarium was different from those of the other six algae at the last two residues （CL to LF）, thus the spatial structure of its GAPDH active center may be different from those of the other six. Phylogenetic analysis indicated that GAPDH of G. lemaneiformis might have undergone an evolution similar to those of Porphyra yezoensis, Chondrus crispus, and Gracilaria verrucosa. C. caldarium had a closer evolutionary relationship with Cyanidioschyzon merolae than with Cyanidium sp. Virtual Northern blot analysis revealed that gpd of G. lemaneiformis expressed constitutively, which suggested that it might be house-keeping and could be adapted as an inner control in gene expression analysis of G. lemaneiformis.     

Automatic cell object extraction of red tide algae in microscopic images

Extracting the cell objects of red tide algae is the most important step in the construction of an automatic microscopic image recognition system for harmful algal blooms.This paper describes a set of composite methods for the automatic segmentation of cells of red tide algae from microscopic images.Depending on the existence of setae,we classify the common marine red tide algae into non-setae algae species and Chaetoceros,and design segmentation strategies for these two categories according to their morphological characteristics.In view of the varied forms and fuzzy edges of non-setae algae,we propose a new multi-scale detection algorithm for algal cell regions based on border-correlation,and further combine this with morphological operations and an improved GrabCut algorithm to segment single-cell and multicell objects.In this process,similarity detection is introduced to eliminate the pseudo cellular regions.For Chaetoceros,owing to the weak grayscale information of their setae and the low contrast between the setae and background,we propose a cell extraction method based on a gray surface orientation angle model.This method constructs a gray surface vector model,and executes the gray mapping of the orientation angles.The obtained gray values are then reconstructed and linearly stretched.Finally,appropriate morphological processing is conducted to preserve the orientation information and tiny features of the setae.Experimental results demonstrate that the proposed methods can effectively remove noise and accurately extract both categories of algae cell objects possessing a complete shape,regular contour,and clear edge.Compared with other advanced segmentation techniques,our methods are more robust when considering images with different appearances and achieve more satisfactory segmentation effects.