Linear and circular dichroism of membranes from Rhodopseudomonas capsulata

Absorption, linear dichroism and circular dichroism spectra of Rhodopseudomonas capsulata (wild-type-St. Louis strain, mutant Y5 and mutant Ala⁺) are particularly sensitive to the nature of the light-harvesting bacteriochlorophyll-carotenoid-protein complexes. Evidence for exciton-type interactions is seen near 855 nm in the membranes from the wild-type and from mutant Y5, as well as in an isolated B-800 + 850 light-harvesting complex from mutant Y5. The strong circular dichroism that reflects these interactions is attenuated more than 10-fold in membranes from the Ala⁺ mutant, which lacks both B-800 + 850 and colored carotenoids and contains only the B-875 light-harvesting complex. These results lead to the conclusion that these two light-harvesting complexes have significantly different chromophore arrangements or local environments.     

An improved method of measuring photosynthetic electron transport in Euglena under in vivo conditions

A method is described to measure photochemical activity in intact cells of Euglena under in vivo conditions. The method employs a cell wall digesting enzyme (cellulysin) to induce enough permeability in the cell walls and membranes in order to allow dyes, commonly used to investigate light-dependent electron transport reactions to enter, but without inducing a concomittant efflux of metabolites. Between 1 and 2 h of incubation in 5% (w/v) cellulysin provided conditions which allowed measurement of photosystem I-, II- and I+II-dependent electron transport with rates up to 600% higher than in control cells; whereas other cell wall degrading enzymes (cellulase and pectinase) still did not increase the entry of the dyes. Cellulysin up to 2 h of incubation had little or no effect on whole cell respiration, photosynthetic O₂ evolution, or the export of potassium and (¹⁴C) labeled compounds out of cells; therefore cellulysin obviously did not change the normal habit or physiology of Euglena. Cellulysin (4 h digestion), cellulase and pectinase (2–4 h of incubation) on the other hand led to a lowering of respiration and light-dependent O₂ evolution, and increased the efflux of K⁺, but apparently decreased that of (¹⁴C)labeled fixation products.Abbreviations DBMIB dibromothymoquinone - DCPIP 2,6-dichlorophenol-indophenol - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DMMIB 2,3-dimethyl-5,6-methylenedioxy-p-benzoquinone - MV methylviologen - PSI photosystem I - PS II photosystem II     

The effects of light quality on the growth of phototrophic bacteria in lakes

The effects of light quality (color) on the ¹⁴CO₂ fixation rates of natural population of photosynthetic sulfur bacteria were tested. The phototrophic bacteria were collected from the sulfide containing waters of 3 stratified lakes. The populations sampled survive in environments where light intensities are very low. Not only are the light intensities low but, due to the light filtering characteristics of the lake water, the light is of specific color. It was determined that the spectral properties of the three lakes differed, hence the quality of light reaching the phototrophic bacteria in each lake differed. It was also observed that only green sulfur bacteria were present in the study lake which transmits mainly red light and both purple and green sulfur bacteria were present in the two study lakes which transmit predominantly green light. Enrichment cultures were set up with phototrophic bacteria from two of the lakes serving as the inocula. Enrichment culture studies and photosynthetic responses of the natural populations indicate that light quality is a major factor in determining the composition of phototrophic bacterial population in some lakes.Non-Common Abbreviations Bchl Bacteriochlorophyll - DPM Disintegrations per minute     

Characterization of the plasmid DNAs of Rhodopseudomonas capsulata

Presence of extrachromosomal DNa in Rhodopseudomonas capsulata strain BH9 was shown by the appearance of a satellite band in a dye-buoyant density gradient. Radioactively labelled DNA was prepared from this satellite band and examined on a 5–20% sucrose gradient. Three radioactive peaks with sedimentation coefficients of 100 S, 94 S, and 58–64 S, respectively, were consistently observed. Analysis of these sedimentation coefficients suggested that there are two species of plasmid DNA with molecular sizes of 94×10⁶ daltons (named pBH91) and 74×10⁶ daltons (named pBH92). The 58–64 S peak is attributed to open circular molecules. DNAs from each peak of the sucrose gradient were examined by electronmicroscopy, and the results agree closely with those of the sucrose gradient analysis. Reassociation kinetics of the plasmid DNA was also followed. Addition of total DNA of strain BH9 increased the renaturation rate of the plasmid DNA. It was calculated from the magnitude of the increase that approximately 10% of the BH9 total DNA may hybridize with the plasmid sequences. DNA prepared from the gene transfer agent (GTA) produced by R. capsulata increases the renaturation rate of the plasmid to the same extent as total DNA isolated from the GTA producing strain, Y262.     

The early formation of the photosynthetic apparatus in Rhodospirillum rubrum

The time dependent assembly of the photosynthetic apparatus was studied in Rhodospirillum rubrum after transfer of cells growing aerobically in the dark to low aeration. While bacteriochlorophyll (Bchl) cellular levels increase continuously levels of soluble cytochrome c ₂do not change significantly. Absorption spectra of membranes isolated at different times after transfer reveal that incorporation of carotenoids lags behind incorporation of Bchl. However, a carotenoid fraction exhibiting spectral properties of spirilloxanthin isomers was isolated apart from membranes. This carotenoid fraction even was present in homogenates from Bchl-free, aerobically grown cells. Incorporation of U-¹⁴C-proteinhydrolyzate into membrane proteins showed that proteins are mainly formed which are specific for photosynthetic membranes. Although the proportion of reaction center (RC) Bchl per light harvesting (LH) Bchl does not change the proportions of membrane proteins present in RC and LH preparations change initially. But later on the proportions of the different proteins also reach constant values. Concerning proteins characteristic for cytoplasmic membranes a differential incorporation of label can be observed. The data indicate that the photosynthetic apparatus in Rhodospirillum rubrum is assembled through a sequential mechanism.Abbreviations Bchl bacteriochlorophyll - LH light harvesting - RC reaction center - R. Rhodospirillum - R. Rhodopseudomonas     

Structure and composition of intracytoplasmic membranes of Ectothiorhodospira mobilis

The lamellar membrane stacks of Ectothiorhodospira mobilis were isolated and purified by a combination of lysozyme and osmotic shock treatment, followed by differential and density gradient centrifugation. Preparations of lamellar membranes were enriched at least 2.4-fold in the ratio of bacteriochlorophyll a to protein.Thin-sectioning, negative staining, platinumcarbon shadowing and freeze-etching were used to study the architecture of the membrane units. Both platinum-carbon shadowing and freeze-etching showed the outer surfaces of the isolated lamellar membrane stacks to be relatively smooth. Particles averaging 7 nm in diameter were seen on several faces following freeze-ctching.Non-polar amino acids amounted to 60% of the total amino acid composition. Lipids constituted 32% of the membrane dry weight. Phosphatidyl ethanolamine and diphosphatidyl glycerol were the major phospholipids. Fatty acids of 10–15 carbons represented a small fraction of both membrane and whole cell fatty acids. Monoenes constituted 36% of the total membrane fatty acids and 38.4% of the total whole cell fatty acids. The major fatty acids of both whole cells and purified membranes were C16:0, C18:1 and cyclopropane C19:0.     

Dependence of the maximum temperature for growth of Saccharomyces cerevisiae on nutrient concentration

Anacystis nidulans (Synechococcus) was maintained in a medium of low phosphate concentration (0.1 mM) and grew with a normal doubling time of 5 hrs at 30°C. Such cultures ahd a normal pigment composition and alkaline phosphatase was detectable at low specific activities only.The onset of phosphate-limited growth occurred when the phosphate concentration in the medium fell to a value below 4 M (the limit of accurate determination by the assay method used) and resulted in increases in alkaline phosphatase activity, reaching a final 10 to 15 fold increase in specific activity after a period of several hours. Marked changes in the overall pigment composition occurred in this period of growth restriction. The addition of phosphate to such cultures resulted in a halt in synthesis of the enzyme and the restoration of normal pigmentation before growth resumed at the normal rate.Several organic phosphate esters could replace inorganic phosphate for growth and were also hydrolyzed by the partially purified enzyme, but growth rates were characteristically lower and the specific activity only 3 to 4 fold higher than in cultures grown in phosphate excess.Studies with the partially purified enzyme suggested that it differed in some of its properties from other alkaline phosphatases described in the literature.Abbreviations Used pNP pnitrophenol - pNPP pnitrophenylphosphate     

Conversion of two distinct Rhodopseudomonas capsulata isolates to the glycerol-utilizing phenotype

Abstract Previous studies employing a single strain of the photosynthetic bacterium Rhodopseudomonas capsulata have demonstrated that this organism, which normally is not capable of growth on glycerol, can gain glycerol-utilizing ability through mutational events. We have examined other strains of R. capsulata of divergent geographic origin, and have found that they could also be converted to a glycerol-catabolic phenotype. Mutant derivatives were found to express high constitutive levels of glycerophosphate dehydrogenase (GPD) and glycerokinase (GK) which were barely detectable in the parental strains. These results suggest that the ability to utilize glycerol after undergoing a gain of function mutation is a widespread property of R. capsulata .     

五唇兰对PEG模拟的干旱胁迫响应研究

为了解干旱对五唇兰(Phalaenopsis pulcherrima)生长的影响,以聚乙二醇(PEG)溶液模拟干旱胁迫,对其叶片的光合色素、渗透调节物质和非结构碳水化合物(NSC)含量变化进行研究。结果表明,随着PEG浓度增加,五唇兰植株含水量和鲜质量逐渐下降,以PEG为13.75%～14.84%时最显著。PEG处理显著降低叶片的叶绿素a和b含量。随着植株含水量的降低,叶片可溶性蛋白、淀粉(St)含量均呈下降趋势,可溶性糖(SS)含量、NSC和SS/St均呈先升后降的趋势。因此,干旱胁迫会影响五唇兰植株的含水量和光合产物的积累;在较低程度干旱胁迫下,可溶性糖在抗旱响应中发挥主要作用;随着干旱胁迫程度加深,五唇兰的生理代谢受到严重影响。     

竹柏2种颜色叶片的光合特性研究

为了解竹柏(Podocarpus nagi)的光合特性,以3 a生全绿叶和花叶竹柏为材料,测定其光合色素含量和气体交换参数。结果表明,全绿叶竹柏叶片的叶绿素a、叶绿素b、类胡萝卜素、叶绿素a+b、叶绿素a/b和叶绿素a+b/类胡萝卜素均显著高于花叶竹柏;全绿叶竹柏叶片的初始量子效率、最大光合速率和暗呼吸速率均显著高于花叶,而光饱和点和光补偿点均显著低于花叶;全绿叶竹柏叶片的初始羧化效率、光合速率、CO2饱和点和光呼吸速率均高于花叶,而CO2补偿点低于花叶。2种颜色叶片的气孔导度、蒸腾速率和水分利用效率均随着光合有效辐射的增大而增大,且均表现为全绿叶花叶,而胞间CO2浓度则相反,表现为花叶全绿叶。因此,全绿叶竹柏利用弱光的能力强于花叶竹柏,而花叶竹柏利用强光的能力更强,在园林绿化配置中,可根据2种颜色叶片的光合特性合理配置。