Dynamics and diffusion in photosynthetic membranes from rhodospirillum photometricum

Photosynthetic organisms drive their metabolism by converting light energy into an electrochemical gradient with high efficiency. This conversion depends on the diffusion of quinones within the membrane. In purple photosynthetic bacteria, quinones reduced by the reaction center (RC) diffuse to the cytochrome bc(1) complex and then return once reoxidized to the RC. In Rhodospirillum photometricum the RC-containing core complexes are found in a disordered molecular environment, with fixed light-harvesting complex/core complex ratio but without a fixed architecture, whereas additional light-harvesting complexes synthesized under low-light conditions pack into large paracrystalline antenna domains. Here, we have analyzed, using time-lapse atomic force microscopy, the dynamics of the protein complexes in the different membrane domains and find that the disordered regions are dynamic whereas ordered antennae domains are static. Based on our observations we propose, and analyze using Monte Carlo simulations, a model for quinone diffusion in photosynthetic membranes. We show that the formation of large static antennae domains may represent a strategy for increasing electron transfer rates between distant complexes within the membrane and thus be important for photosynthetic efficiency.     

Spherical subunits in photosynthetic membranes of two cyanophyceae and the bacterium rhodospirillum rubrum

Summary The photosynthetic lamellae of Oscillatoria amoena and Pseudanabaena catenata appear as a two-dimensional packing of spherical units following permanganate fixation, ultrathin sectioning and high-resolution electron micrography. The usual three-layered appearance is explained as an optical artifact. Quite similar patterns were observed in chromatophore membranes of Rhodospirillum rubrum. Also, the cytoplasmic membrane of R. rubrum shows an identical pattern.Indication is presented that thylakoids of Cyanophyceae can arise as invaginations of the cytoplasmic membrane.The localization of chloroplast lipids in these organisms and the similarity of spherical units and quantasomes is discussed.     

On evolution of the photosynthetic pigments

Some considerations on the origin of the photosynthetic pigment apparatus in the period of transition of organisms from heterotrophic to phototrophic mode of life and further evolution of this apparatus are given in this paper.Some ideas about interrelation between major and auxiliary pigments and about different forms of the pigments in vivo are discussed from an evolutionary point of view.Special Symposium on Photochemistry and the Origins of Life, Sixth International Congress on Photobiology, Bochum, Germany.     

On the evolution of the photosynthetic pigments

During the course of terrestrial evolution, some organisms developed the capability of capturing and utilizing solar radiation. Colored compounds were undoubtedly incorporated within living forms from the earliest times, but during the transition from heterotrophic to a photoautotrophic metabolism only those pigments were selected that were components of the evolving photosynthetic apparatus and were able to catalyze reactions involving storage of light energy in chemical bonds. In this communication, some properties of tetrapyrroles with a closed porphyrin ring containing a metal ion in the center are discussed. These compounds are present in all principal contemporary photosynthetic pigments, and their synthesis has been demonstrated from simpler compounds under prebiotic conditions. It is probable that during intermediate stages in the evolution of photosynthesis, pigments with oxidizing potentials lower than that of chlorophyll were utilized to store light energy although they were not capable of removing electrons from water. The evolution and function of multiple forms of a given photosynthetic pigmentin vivo are discussed. ‘Accessory’ pigments may be regarded as rudiments of the evolutionary development of the photosynthetic apparatus.     

Distribution of phosphatidylethanolamine in the lipid bilayer of chromatophores of the photosynthetic bacterium rhodospirillum rubrum

The distribution of phosphatidylethanolamine in the two lipid layers of chromatophores ofRhodospirillum rubrum has been analysed by chemical modification of phosphatidylethanolamine (PE) with trinitrobenzenesulfonic acid (TNBA) at low temperatures. Around 45±1% of the total phosphatidylethanolamine is labelled by this procedure independent on chromatophore purity, vesicle size, action of proteases and growth state of the cells. This demonstrates a complete modification of the accessible phosphatidylethanolamine and an asymmetric distribution of phosphatidylethanolamine, with 45% of the phosphatidylethanolamine in the outer part of the bilayer.Abbreviations TNBA 2,4,6 trinitrobenzenesulfonic acid - PE phosphatidylethanolamine - PMS phenazinmethosulfate     

On the evolution of the photosynthetic pigments.

During the course of terrestrial evolution, some organisms developed the capability of capturing and utilizing solar radiation. Colored compounds were undoubtedly incorporated within living forms from the earliest times, but during the transition from heterotrophic to a photoautotrophic metabolism only those pigments were selected that were components of the evolving photosynthetic apparatus and were able to catalyze reactions involving storage of light energy in chemical bonds. In this communication, some properties of tetrapyrroles with a closed porphyrin ring containing a metal ion in the center are discussed. These compounds are present in all principal contemporary photosynthetic pigments, and their synthesis has been demonstrated from simpler compounds under prebiotic conditions. It is probable that during intermediate stages in the evolution of photosynthesis, pigments with oxidizing potentials lower than that of chlorophyll were utilized to store light energy although they were not capable of removing electrons from water. The evolution and function of multiple forms of a given photosynthetic pigment in vivo are discussed. 'Accessory' pigments may be regarded as rudiments of the evolutionary development of the photosynthetic apparatus.     

Bio-optical modeling of photosynthetic pigments in corals

The spectral reflectance of coral is inherently related to the amounts of photosynthetic pigments present in the zooxanthellae. There are no studies, however, showing that the suite of major photosynthetic pigments can be predicted from optical reflectance spectra. In this study, we measured cm-scale in vivo and in situ spectral reflectance for several colonies of the massive corals Porites lobata and Porites lutea, two colonies of the branching coral Porites compressa, and one colony of the encrusting coral Montipora flabellata in Kaneohe Bay, Oahu, Hawaii. For each reflectance spectrum, we collected a tissue sample and utilized high-performance liquid chromatography to quantify six major photosynthetic pigments, located in the zooxanthellae. We used multivariate multiple regression analysis with cross-validation to build and test an empirical linear model for predicting pigment concentrations from optical reflectance spectra. The model accurately predicted concentrations of chlorophyll a, chlorophyll c ₂, peridinin, diadinoxanthin, diatoxanthin and β-carotene, with correlation coefficients of 0.997, 0.941, 0.995, 0.996, 0.980 and 0.984, respectively. The relationship between predicted and actual concentrations was 1:1 for each pigment, except chlorophyll c ₂. This simple empirical model demonstrates the potential for routine, rapid, non-invasive monitoring of coral-zooxanthellae status, and ultimately for remote sensing of reef biogeochemical processes.     

A TLC-GLC procedure for the quantitation of algal photosynthetic pigments and their degradation products

A procedure employing and GLC techniques for the analysis of algal chlorophylls and their degradation products has been developed and evaluated. Algal pigments were separated on MN 300 cellulose plates developed in an ascending solvent system of hexane saturated with acetonitrile and n-propanol (100:0,4, v:v). Quantitation of chlorophyll a, b and pheophytin a, b were accomplished by GLC analysis of their phytol, following alkaline methanolic hydrolysis of the individual pigments. The amount of chlorophyllides/pheophorbides in a sample was estimated by its free phytol content. This technique is especially valuable for the evaluation of the pigment contents of near sediment phytoplankton and periphyton samples, where large quantities of chlorophyll degradation products and/or carotenoid pigments are generally present which may interfere significantly with the routine analytical methods.     

Cooperative polymerization of photosynthetic pigments in formamide-water solution

The aggregation of bacteriochlorophyll a and bacteriopheophytin a into large oligomers with maximum optical absorption at 860 nm was studied in a 3:1 (vol/vol) formamide/water solution, using optical absorption spectroscopy and electron microscopy. The aggregation is cooperative and proceeds according to two equilibrium constants. Initially, two pigment molecules form a “seed” that absorbs at ≈860 nm. The equilibrium constant, K_a, governing this reaction equals 1.3 × 10³ M^-1 in the case of bacteriochlorophyll a (due to experimental limitations, K_a for bacteriopheophytin a could not be determined). The addition of monomers to aggregates consisting of two or more units is governed by an equilibrium constant, K_b, equal to 2.2 × 10⁶ M^-1 for bacteriochlorophyll a and ≈ 10⁹ M^-1 for bacteriopheophytin a. The enthalpy and entropy changes that drive the bacteriochlorophyll oligomer formation are -9.25 and ≈0.0 kcal/mol, respectively. Above a threshold concentration, the amount of oligomers remains constant but their length continues to increase. Each oligomer appears to consist of dimers that are associated by hydrophobic interactions among their alcohol residues, forming long strands. Single strands presumably coil into helices that are seen as cylinders. The bacteriochlorophyll a oligomers form cylinders with a constant diameter of 150 Å and an average length of 2,000 Å (at 1.5 × 10^-5 M bacteriochlorophyll a). These cylinders contain 200-250 bacteriochlorophyll a dimers. The bacteriopheophytin oligomers coil into wider cylinders (≈400 Å in diameter) which contain ≈600-700 bacteriopheophytin a dimers. In both cases, the separation between the dimers is ≈20 Å. At such distances, the dipolar interactions among adjacent dimers are negligible and do not affect the optical absorption of each individual pair. Therefore, the optical absorption of these pairs can be a tool for investigating the absorption pattern of photosynthetic pigments in vivo.     

Genes involved in the biosynthesis of photosynthetic pigments in the purple sulfur photosynthetic bacterium Thiocapsa roseopersicina

A pigment mutant strain of the purple sulfur photosynthetic bacterium Thiocapsa roseopersicina BBS was isolated by plasposon mutagenesis. Nineteen open reading frame, most of which are thought to be genes involved in the biosynthesis of carotenoids, bacteriochlorophyll, and the photosynthetic reaction center, were identified surrounding the plasposon in a 22-kb-long chromosomal locus. The general arrangement of the photosynthetic genes was similar to that in other purple photosynthetic bacteria; however, the locations of a few genes occurring in this region were unusual. Most of the gene products showed the highest similarity to the corresponding proteins in Rubrivivax gelatinosus. The plasposon was inserted into the crtD gene, likely inactivating crtC as well, and the carotenoid composition of the mutant strain corresponded to the aborted spirilloxanthin pathway. Homologous and heterologous complementation experiments indicated a conserved function of CrtC and CrtD in the purple photosynthetic bacteria. The crtDC and crtE genes were shown to be regulated by oxygen, and a role of CrtJ in aerobic repression was suggested.     

分离自我国蜜蜂的一株新的致病螺原体及其基本特性

【目的】通过分析分离自我国患病蜜蜂体内的螺原体MF1006的基本生物学特征,初步确定其分类地位及致病性。【方法】应用暗视野显微镜和透射电子显微镜观察螺原体形态,运用常规螺原体分离和培养方法、分子生物学方法和血清学方法研究螺原体分离菌株可能的分类地位,并采用饲喂接种法研究其致病性。【结果】菌株MF1006具有典型的螺旋形和运动性,能通过0.22μm孔径的滤膜,与我国之前发现的引起蜜蜂螺原体病的参比菌株Spiroplasma melliferum CH-1的基本生物学特征差异较大。S.melliferum CH-1抗血清对其没有抑制作用。根据16S rDNA、ITS序列构建系统发育树显示,菌株MF1006与在法国发现的引起蜜蜂"五月病"的Spiroplasma apis的亲缘关系最近。此外,菌株MF1006对供试意蜂有较强的致病性。【结论】分离菌株MF1006是在我国蜜蜂体内发现的除S.melliferum以外的另一种致病螺原体。     

An improved method for extraction and separation of photosynthetic pigments

The method for extracting and separating hydrophobic photosynthetic pigments proposed by Katayama et al. (Japanese Journal of Phycology, 42, 71-77, 1994) has been improved to introduce it to student laboratories at the senior high school level. Silica gel powder was used for removing water from fresh materials prior to extracting pigments by a mixture of organic solvents that was also used for chromatographic separation of the pigments. A small silica gel thin-layer plate or a paper strip was used for separating the pigments. The improved method may be applicable to all kinds of plant materials including algae, is easier than most other methods, and can lead to more successful results in separating these pigments by both thin-layer chromatography and paper chromatography. The method has been carried out in student laboratories in some senior high schools and universities in Japan. The results indicate that this laboratory exercise is effective for students to recognise the unity and diversity of plants. Therefore, this laboratory seems to be useful for teaching plant systematics as well as for teaching photosynthesis.     

Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments

Photopigment radiolabcling, a useful method for measuring thein situ carbon-specific growth rates of microalgae, is basedon the determination of synthesis rates of chemosystematic (i.e.specific for microalgal phylogenetic groups) chlorophylls andcarotenoids using photosynthetically assimilated ¹⁴C as a radiotracer.The reliability of this method depends on accurate measurementsof the ¹⁴C-specific activity of individual photopigments. Typically,photopigments are separated by high-performance liquid chromatography(HPLC) with fraction collection of individual peaks, followedby further purification and standard scintillation counting.To simplify analyses, we evaluated in-line flow scintillationcounting to determine its applicability and reliability formeasuring the activity of radio-labeled photopigments. Incubationswere conducted using both pure cultures and natural phyto-planktonsamples. The radiochemical purity of photopigments was determinedby extract acidification (10% HC1) to transform chlorophyllsinto degradation products. Purity was also checked by comparingabsorbance spectra with purified standards. Although ¹⁴C-labeledcolorless compounds are a common feature in radiograms, thesecompounds do not co-elute with photopigments using our HPLCprotocol. Flow scintillation counting, coupled with a highlyselective HPLC protocol, provides an efficient, reliable andfeasible alternative to fraction collection/repurification methodsfor measuring the ¹⁴C-specific activity of microalgal photosyntheticpigments.     

Interactions between photosynthetic pigments in organisms and in model systems

The review summarizes results concerning photosynthetic systems with chlorophylls and carotenoids obtained by means of spectral methods such as polarized radiation, photoacoustic spectroscopy, delayed luminescence, thermal deactivation, and leading to construction of model systems.