三种紫球藻亲缘关系RAPD分析

利用RAPD技术对铜绿紫球藻（Porphyridium aerugineum 755）,淡色紫球藻（Porphyridium purpureum 806）和紫球藻（Porphyridium cruentum）的亲缘关系进行分析。从50个随机引物中筛选出25个种间多态性较强,重复性较好的引物。检测到233个位点。在233个条带中有186个多态性位点。多态位点比率为79.73%,平均每个引物扩增9个条带,多态性条带7个。聚类分析结果表明：铜绿紫球藻、淡色紫球藻和紫球藻之间的平均遗传距离为0.455,其中铜绿紫球藻和淡色紫球藻之间的遗传距离为0.413,铜绿紫球藻和紫球藻之间的遗传距离为0.556,淡色紫球藻和紫球藻之间的遗传距离最小为0.396。所以由RAPD试验的分析结果可以得出:三种紫球藻为独立的种, 其中淡色紫球藻和紫球藻是亲缘关系较近的两个不同种。紫球藻属种间个体DNA多态性比较丰富,因此利用RAPD技术可以从DNA水平上检测紫球藻属种间差异。     

Energy transfer from photosystem II to photosystem I in Porphyridium cruentum

Rates of photooxidation of P-700 by green (560 nm) or blue (438 nm) light were measured in whole cells of Porphyridium cruentum which had been frozen to ?196 °C under conditions in which the Photosystem II reaction centers were either all open (dark adapted cells) or all closed (preilluminated cells). The rate of photooxidation of P-700 at ?196 °C by green actinic light was approx. 80% faster in the preilluminated cells than in the dark-adapted cells. With blue actinic light, the rates of P-700 photooxidation in the dark-adapted and preilluminated cells were not significantly different. These results are in excellent agreement with predictions based on our previous estimates of energy distribution in the photosynthetic apparatus of Porphyridium cruentum including the yield of energy transfer from Photosystem II to Photosystem I determined from low temperature fluorescence measurements.     

Primary Events, Energy Transfer, and Reactions in Photosynthetic Units: Lifetime of the Excited State In Vivo: I. Chlorophyll a in Algae, at Room and at Liquid Nitrogen Temperatures; Rate Constants of Radiationless Deactivation and Trapping

Using a mode-locked laser (λ, 632.8 nm), fluorescence decay of chlorophyll (Chl) a in the green alga Chlorella pyrenoidosa, the red alga Porphyridium cruentum, and the blue-green alga Anacystis nidulans was measured by the phase-shift method under conditions when photosynthesis was not operative (3-(3,4-dichlorophenyl)-1,1-dimethylurea [DCMU] poisoning, or cooling to 77°K). In the presence of 10^-5 M DCMU, the lifetime of Chl a fluorescence (τ) at room temperature is about 1.7 nsec in Chlorella, 1.0 nsec in Porphyridium, and 0.7 nsec in Anacystis. At 77°K, τ is 1.4 nsec (for fluorescence at about 685 nm, F-685) and 2.3 nsec (for F-730) in Chlorella, 0.9 nsec (F-685) and 1.2 nsec (F-730) in Porphyridium, and 0.8 nsec (F-685 and F-730) in Anacystis. From the above measurement, and the assumption that τ₀ (the intrinsic fluorescence lifetime) for Chl a in all three algae is 15.2 nsec, we have calculated the rate constants of radiationless transition (that includes energy transfer to weakly fluorescent system I) processes competing with fluorescence at room temperature to be about 5 × 10⁸ sec^-1 in Chlorella, 9 × 10⁸ sec^-1 in Porphyridium, and 13 × 10⁸ sec^-1 in Anacystis. At 77°K, this rate constant for Chl a that fluoresces at 685 nm remains, in the first approximation, the same as at room temperature. From the τ data, the rate constant for the trapping of excitation energy is calculated to be about 1.2 × 10⁹ sec^-1 for Chlorella, 2 × 10⁹ sec^-1 for Porphyridium, and 2 × 10⁹ sec^-1 for Anacystis. The efficiency of trapping is calculated to be about 66% (Chlorella), 68% (Porphyridium), and 60% (Anacystis). (It is recognized that variations in the above values are to be expected if algae grown under different conditions are used for experimentation.) The maximum quantum yield of Chl a fluorescence for system II (λ, 632.8 nm), calculated from τ measurements, is about 10% in Chlorella, 6-7% in Porhyridium, and 5% in Anacystis under conditions when photosynthesis is not operative; the values at 77°K appear to be very close to those with DCMU added at room temperature. ø for F-730 at 77°K, however, is somewhat higher than for F-685. The predicted quantum yields of fluorescence for Chl a in intact cells (both systems I and II) at low intensities of 632.8 nm light are about 2-3, 1-2, and 1% for Chlorella, Porphyridium, and Anacystis, respectively.     

Mixotrophic production of phycoerythrin and exopolysaccharide by the microalga Porphyridium cruentum

The ability of the unicellular rhodophyte Porphyridium cruentum to grow mixotrophically on the soluble fraction of Solarium tuberosum meal was tested. At the beginning of stationary phase Porphyridium cruentum produced 7 μg ml⁻¹ of phycoerythrin and 129 μg ml⁻¹ of total soluble exopolysaccharide when cultured autotrophically. When cultured mixotrophically with the soluble fraction of Solanum tuberosum meal, the productivity increased to 10 μg ml⁻¹ of phycoerythrin and 330 μg ml⁻¹ of total soluble exopolysaccharide. When the soluble fraction of S. tuberosum meal was supplied together with nitrate and phosphate, the productivity of phycoerythrin increased to 21 μg ml⁻¹ while the production of total soluble exopolysaccharide decreased to 195 μg ml⁻¹. Results demonstrate that the soluble fraction of S. tuberosum meal can be used as substrate for the production of phycoerythrin and exopolysaccharide by P. cruentum improving the results obtained with the autotrophic culture medium.     

Porphyridium violaceum,eine marine neue Art

Zusammenfassung 1. Die neubeschriebene marinePorphyridium-Art unterscheidet sich von dem terrestrisch lebenden, aber auch im Meerwasser gefundenenPorphyridium purpureum durch ihre größeren, violett gefärbten Zellen.2. Teilung und Wachstum der Zellen vonPorphyridium violaceum passen sich dem Belichtungsrhythmus an. Die während der 10stündigen Dunkelheit geteilten Zellen sind 9 bis 11µ dick, sie vergrößern sich während der Lichtperiode auf 11 bis 14µ.

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Porphyridium violaceum, a marine new species

The new species is marine, whereasPorphyridium purpureum (= P. cruentum), first known only from terrestrial habitats, is halophilic and has been found several times in seawater.Porphyridium violaceum differs from the above mentioned species by a larger diameter and the colour of its cells.

     

Effects of light intensity on the growth rate of the red alga Porphyridium cruentum

The red alga, Porphyridium cruentum, which is one of the potential sources of arachidonic acid, was cultured in batch and continuous vessels. The growth rates in batch cultures were correlated to the mean light intensity in the vessels, and the cell concentrations in continuous cultures were estimated by those results. The yield of arachidonic acid was about 1.2 g per 10¹² cell at cell concentrations ranging from 0.5 to 1.5 × 10¹⁰ cell/l and independent of the mean light intensity.     

Thermal and pH Stability of the B-Phycoerythrin from the Red Algae Porphyridium cruentum

Phycoerythrin is the major light-harvesting pigment-protein of the red algae Porphyridium cruentum and is widely used as fluorescent probe and analytical reagent. Additionally this protein has a potential application as natural dye in food industry. Nevertheless the knowledge of the functional properties of this alga protein is limited, hindering its application as food additive. In this article we report a biophysical characterization of B-phycoerythrin from Porphyridium cruentum (B-PE) in order to study its stability and spectral properties in a broad range of pHs. This information can help in its potential application as colorant in the food industry. Spectroscopic data obtained in this work show that B-PE has a stronger functional stability in the pH range 4.0–10.0, and Size Exclusion Chromatography suggests that the protein maintains a (αβ)₆-γ oligomeric structure in that range of pHs. At pH 7.0, an apparent T _m value of 77.5?±?0.5 °C was calculated. At this pH, the protein is highly stable with a loss of only 20 % of its spectral properties (absorbance and fluorescence) after 25 days at room temperature. These results indicate that B-PE is more stable in a broad range of pHs than other phycoerythrin proteins, which would facilitate its use in the food industry.     

Overproduction of gamma-Linolenic and Eicosapentaenoic Acids by Algae

The pharmaceutical interest and limited availability of γ-linolenic acid (GLA) and eicosapentaenoic acid (EPA) prompted the search for genetic means for increasing the production of these fatty acids from algal sources. Cell lines of Spirulina platensis and Porphyridium cruentum resistant to the growth inhibition of the herbicide Sandoz 9785 were selected by serial transfers of the culture in the presence of increasing concentrations of the herbicide. The resistant cell lines of S. platensis overproduced GLA and those of P. cruentum overproduced EPA and were stable for at least 50 generations in the absence of the inhibitor.     

Indoor mass cultivation of red alga Porphyridium cruentum in different types of bioreactors: effect of scale-up and vessel shape

The present study describes scale-up of Porphyridium cruentum cultures in different types of enclosed bioreactors. Special consideration for the purpose was the comparative performance of these photobioreactors relevant to growth and polysaccharide production, which have a significance in the commercial exploitation of the microalga. The custom-built flat-sided photobioreactor with higher exposed surface area to volume ratio was found to be the best system for the cultivation of P. cruentum.     

N-terminus conservation in the terminal pigment of phycobilisomes from a prokaryotic and eukaryotic alga

High molecular weight polypeptides from phycobilisomes, believed to be involved in facilitating the energy flow from phycobilisomes to thylakoids, are conserved in the prokaryote Nostoc sp. and the eukaryote Porphyridium cruentum. Partial N-terminal sequence analysis of the phycobilisome-polypeptides of Nostoc (94 kilodalton) and Porphyridium (92 kilodalton) revealed 55% identity in the first 20 residues, but no significant homology with sequences of other phycobiliproteins or phycobilisome-linkers. Polypeptides (94 and 92 kilodalton) from Nostoc thylakoids free of phycobilisomes, previously presumed to be involved in the phycobilisome-thylakoid linkage (M Mimuro, CA Lipschultz, E Gantt 1986 Biochim Biophys Acta 852: 126) exhibit the same immunocrossreactivity but are different from the 94 kilodalton-phycobilisome polypeptide by having blocked N-termini and a different amino acid composition.     

Effective Absorption Cross-Sections in Porphyridium cruentum: Implications for Energy Transfer between Phycobilisomes and Photosystem II Reaction Centers

Effective absorption cross-sections for O₂ production by Porphyridium cruentum were measured at 546 and 596 nanometers. Although all photosystem II reaction centers are energetically coupled to phycobilisomes, any single phycobilisome acts as antenna for several photosystem II reaction centers. The cross-section measured in state I was 50% larger than that measured in state II.     

Preparation of different molecular weight polysaccharides from Porphyridium cruentum and their antioxidant activities

Hermetical microwave was used to degrade Porphyridium cruentum polysaccharides from 2918 to 256.2, 60.66 and 6.55 kDa. The antioxidant properties of different molecular weight polysaccharides were evaluated by determining the scavenging ability of free radicals, inhibitory effects on lipid peroxidation in liver homogenates and hemolysis of mouse erythrocytes. Analysis of physicochemical properties confirmed that microwave degradation might not markedly change the chemical components of the polysaccharides. High-molecular-weight polysaccharides from P. cruentum had no obvious antioxidant activity, but low-molecular-weight fragments after degradation exerted an inhibitory effect on oxidative damage. The 6.55-kDa fragment had stronger antioxidant activity than the 60.66 and 256-kDa fragments.     

In vivo molybdate inhibition of sulfate transfer to porphyridium capsular polysaccharide

Active transport of exogenous sulfate into log phase cells of Porphyridium aerueineum followed Michaelis-Menten kinetics, and the apparent Km for sulfate transport is approximately 2.5 × 10⁻⁶m. Molybdate, also a group VI anion, is a competitive inhibitor of sulfate transport, and the inhibition is freely reversible. Once in the cell, molybdate depresses the rate of sulfate pool utilization by blocking sulfate transfer to polysaccharides destined for secretion to the cell surface. Specifically, molybdate inhibits the formation of adenosine 5′-phosphosulfate and in turn the formation of adenosine 3′-phosphate 5′-phosphosulfate, the activated donor for sulfate transfer reactions. Combined with the previous identification of adenosine 3′-phosphate 5′-phosphosulfate, this is taken as evidence that the adenosine 5′-phosphosulfate/adenosine 3′-phosphate 5′-phosphosulfate enzymatic sequence for sulfate activation and sulfate donor reactions is operating in Porphyridium. Thiosulfate is utilized as effectively as sulfate as both a sulfur source for growth and polysaccharide synthesis.     

Low molecular weight carbohydrates in Cyanidium caldarium and some related algae

Floridoside (2-O-glycerol-α-d-galactopyranoside) and a small amount of iso-floridoside (1-O-glycerol-α-d- galactopyranoside) were found in Cyanidium caldarium. Floridoside was also found in the red algae Porphyridium cruentum and Porphyra yezoensis, although in the latter iso-floridoside was the main component. Sucrose and glucose were found in the green algae Chlorella pyrenoidosa and Scenedesmus obliquus, and also in a blue-green alga, Anacystis nidulans. Another blue-green alga, Phormidium foveolarum, contains mostly trehalose. From these results and from morphological considerations, it is suggested that Cyanidium caldarium belongs to the primitive Rhodophyta.     

The development of Porphyridium purpureum (Bory de Saint-Vincent) Drew et Ross, 1965 (Rhodophyta) from Amursky Bay,Sea of Japan,in a laboratory culture

The growth of the microscopic red alga Porphyridium purpureum was experimentally studied in different culture media and at various levels of water salinity. The ability of this species to recover after cryopreservation was analyzed. It was revealed that the growth rate and generation time of P. purpureum during the exponential growth phase did not significantly differ between cultures on f medium and those on Goldberg’s medium. The species has been found to withstand salinity variations ranging from 8 to 32 vol %. Cryopreservation of P. purpureum cells using three-step freezing with trehalose as a cryoprotectant showed that the dynamics of cell density after thawing insignificantly differed from the control ones.     

Inhibition of Photosynthesis in Certain Algae by Extreme Red Light

This paper shows that in Porphyridium cruentum and in Chlorella pyrenoidosa (but apparently not in Anacystis nidulans) “extreme red” light (> 720 mμ) can inhibit photosynthesis produced by “far red” light (up to 720 mμ). From the action spectrum of this phenomenon, it appears that an unknown pigment with an absorption band around 745 mμ must be responsible for it.     

Photosynthetic Light Reactions in Chemically Fixed Anacystis nidulans, Chlorella pyrenoidosa, and Porphyridium cruentum

The photochemical activities of various species of unicellular algae (Anacystis nidulans, Chlorella pyrenoidosa, and Porphyridium cruentum) were studied following chemical fixation. Fixation with formaldehyde and glutaraldehyde yielded cells which retained their ability to perform photosystem I and photosystem II reactions. The photochemical efficiencies of some fixed algae are as great as those of unfixed spinach chloroplasts. Fixed algae containing accessory pigments appear to be useful models for further studies of the light reactions of photosynthesis.     

Selenium containing amino acids and proteins in marine algae

The unicellular marine algae, Dunaliella primolecta Butcher, Chlorella sp. and Porphyridium cruentum (S.F. Grey) were grown in artificial sea water containing a sublethal concentration of selenite, 10^?2 g Se/1. Both free-and protein-bound seleno-amino acids were identified. The initial steps of selenium incorporation seem to involve the use of the sulfur enzymatic machinery resulting in the replacement of some of the sulfur by selenium in both free amino acids and proteins. At relatively low selenium concentrations, selenium-specific enzymes seem to be in operation.     

The quaternary structure of a unique phycobiliprotein: B-phycoerythrin from Porphyridium cruentum

B-phycoerythrin, from the unicellular red alga Porphyridium cruentum, was crystallized in the rhombohedral space group R3 with a=111.0Å and α=116.8° or A=B=189.1Å and C=60.1Å and γ=120°. Density measurements on the crystals indicate that the hexagonal unit cell can acconmodate three cylindrical molecules, 109Å in diameter and 60Å in height, each of approximately 275,000 daltons. The crystallographic symmetry of the unit cell requires at least 3-fold symmetry for the particle. However, the particle stoichiometry has been reported as (αβ)₆γ and this composition is also supported by SDS gel electrophoresis on the crystalline protein. These results are discussed in light of preliminary model calculations on the quaternary structure of B-phycoerythrin.     

Isolation and Function of Allophycocyanin B of Porphyridium cruentum

Allophycocyanin B was purified to homogeneity from the eukaryotic red alga Porphyridium cruentum. This biliprotein is distinct from the allophycocyanin of P. cruentum with respect to subunit molecular weights, and spectroscopic and immunological properties. The purified allophycocyanin B has a long wavelength absorption maximum at 669 nm at room temperature and at 675 nm at −196 C while the fluorescence emission maximum is at 673 nm at room temperature and 679 nm at −196 C. The emission spectrum of allophycocyanin shifted only 1 nm, from 659 to 660 nm, on cooling to −196 C, and was the same with allophycocyanin crystals as it was with pure solutions of the pigment. Phycobilisomes from P. cruentum have a major fluorescence emission band at 680 nm at −196 C which emanates from the small amount of allophycocyanin B present in the phycobilisomes. Light energy absorbed by the bulk of the biliprotein pigments is transferred to allophycocyanin B with high efficiency.