Photosynthetic action spectra and adaptation to spectral light distribution in a benthic cyanobacterial mat.

We studied adaptation to spectral light distribution in undisturbed benthic communities of cyanobacterial mats growing in hypersaline ponds at Guerrero Negro, Baja California, Mexico. Microscale measurements of oxygen photosynthesis and action spectra were performed with microelectrodes; spectral radiance was measured with fiber-optic microprobes. The spatial resolution of all measurements was 0.1 mm, and the spectral resolution was 10 to 15 nm. Light attenuation spectra showed absorption predominantly by chlorophyll a (Chl a) (430 and 670 nm), phycocyanin (620 nm), and carotenoids (440 to 500 nm). Blue light (450 nm) was attenuated 10-fold more strongly than red light (600 nm). The action spectra of the surface film of diatoms accordingly showed activity over the whole spectrum, with maxima for Chl a and carotenoids. The underlying dense Microcoleus population showed almost exclusively activity dependent upon light harvesting by phycobilins at 550 to 660 nm. Maximum activity was at 580 and 650 nm, indicating absorption by phycoerythrin and phycocyanin as well as by allophycocyanin. Very little Chl a-dependent activity could be detected in the cyanobacterial action spectrum, even with additional 600-nm light to excite photosystem II. The depth distribution of photosynthesis showed detectable activity down to a depth of 0.8 to 2.5 mm, where the downwelling radiant flux at 600 nm was reduced to 0.2 to 0.6% of the surface flux.     

Photosynthetic action spectra and adaptation to spectral light distribution in a benthic cyanobacterial mat

We studied adaptation to spectral light distribution in undisturbed benthic communities of cyanobacterial mats growing in hypersaline ponds at Guerrero Negro, Baja California, Mexico. Microscale measurements of oxygen photosynthesis and action spectra were performed with microelectrodes; spectral radiance was measured with fiber-optic microprobes. The spatial resolution of all measurements was 0.1 mm, and the spectral resolution was 10 to 15 nm. Light attenuation spectra showed absorption predominantly by chlorophyll a (Chl a) (430 and 670 nm), phycocyanin (620 nm), and carotenoids (440 to 500 nm). Blue light (450 nm) was attenuated 10-fold more strongly than red light (600 nm). The action spectra of the surface film of diatoms accordingly showed activity over the whole spectrum, with maxima for Chl a and carotenoids. The underlying dense Microcoleus population showed almost exclusively activity dependent upon light harvesting by phycobilins at 550 to 660 nm. Maximum activity was at 580 and 650 nm, indicating absorption by phycoerythrin and phycocyanin as well as by allophycocyanin. Very little Chl a-dependent activity could be detected in the cyanobacterial action spectrum, even with additional 600-nm light to excite photosystem II. The depth distribution of photosynthesis showed detectable activity down to a depth of 0.8 to 2.5 mm, where the downwelling radiant flux at 600 nm was reduced to 0.2 to 0.6% of the surface flux.     

Protochlorophyll forms with different molecular arrangements

Spectral properties of protochlorophyll (PChl) forms were investigated in solid-film model systems by absorption. fluorescence and circular dichroism (CD) spectroscopy. The solid films were prepared from diethyl ether solution of PChl on a cover glass surface by evaporation of the solvent. After preparation the films usually showed an absorption maximum at 635 nm or in some cases at 640 nm. The PChl form with 635 nm absorption maximum had no CD signal, whilst the films with absorption maximum at 640 nm gave an intense negative CD band at about 640 nm and a positive one at 668 nm. The treatment of the films with ammonia or acetone vapour resulted in a red shift of the absorption maximum from 635 nm or 640 nm to 650 nm. The study of the CD spectra of the films with different PChl forms showed that, depending on the treatment, forms of PChl with similar absorption and fluorescence spectra, but with opposite CD signals, can exist. It is suggested that the differences of the CD spectra are mainly due to different arrangements of the aggregates.     

Wavelength dependence of inactivation and membrane damage to Saccharomyces cerevisiae cells by monochromatic synchrotron vacuum-uv radiation (145-190 nm)

Using an electron storage ring as a source of radiation, the wavelength dependence of inactivation and membrane damage in yeast cells (Saccharomyces cerevisiae) was investigated in the range from 145 to 254 nm, with special reference to the effects of vacuum-uv radiation. The cells were irradiated on a Millipore filter in a moist chamber filled with water vapor (deoxygenated) at saturation pressure. Fluence-survival curves taken at 5-nm intervals were generally sigmoidal. Action spectra of the two types of effects were nearly identical in shape. The maximum occurred in both spectra at 160 nm, decreasing sharply toward 180 nm. The spectra remarkably resembled the calculated absorption spectrum of (liquid) water in the range from 145 to 170 nm; the spectra had no similarity at all to the absorption spectra of DNA, proteins, or lipids. These data support the theory that inactivation of wet cells by vacuum-uv radiation may be attributable to damage in the cell membrane initiated by the absorption of water molecules. Above 210 nm the spectrum for inactivation paralleled the absorption of DNA. Genetic changes (induction of gene conversion) were also observed above 210 nm. Photoreversion for the induced convertants was detectable only above 220 nm. These characteristics are consistent with the expectation that above 210 nm the site of major lethal damage shifts to DNA.     

Luminescence study of monodispersed ZnS nanoparticles

Monodispersed ZnS nanoparticles have been successfully synthesized by a chemical precipitation method in an air atmosphere using polyvinylpyrrolidone (PVP) and sodium hexametaphosphate (SHMP) as surfactants. The synthesized nanoparticles were characterized by X‐ray diffraction (XRD), transmission electron microscopy (TEM), Fourier transform infrared spectrometer (FT‐IR), UV–Vis optical absorption and photoluminescence (PL) spectra. Prepared surfactants capped ZnS nanoparticles are highly homogeneous and well dispersed. Optical absorption spectra showed a strong blue shift from the uncapped particles due to the quantum confinement effect. The capped ZnS emission intensity is enhanced than more the uncapped particles. The size of the synthesized particles is around 4–6.5 nm range. Copyright © 2012 John Wiley & Sons, Ltd.     

Synthesis, self-assembly and characterization of a new glucoside-type hydrogel having a Schiff base on the aglycon

A new hydrogel based on a substituted phenyl glucoside with a Schiff base in the aglycon was synthesized, and the self-assembling characteristics was studied. FTIR spectra, UV-vis absorption spectra and X-ray diffraction (XRD) revealed that π-π interactions between the Schiff base moieties, hydrogen bonds, and the interdigitated interactions between hydrophobic chains had effects on the formation of the self-assembling hydrogel. Scanning electron microscopic (SEM) and transmission electron microscopic (TEM) observation showed that the three-dimensional hydrogel network was constructed from nanotubes with inner diameters of ca. 75 nm and wall of ca. 20 nm.     

Spectral characteristics of rice plants infested by brown planthoppers

Spectral characteristics of rice plants at various levels of infestation by the brown planthopper, Nilaparvata lugens (St?l), (Homoptera:Delphacidae), in the early grain-filling stage were measured and analyzed using a spectroradiometer. Plant damage was classified into six scales, i.e., 0 (CK), 1, 3, 5, 7 and 9, based on the scale of infestation displayed on the surfaces of plant parts. Results showed that mean curves of reflectance spectra (350 - 1800 nm) from different scales of insect infestation were clearly differentiated, especially in the region of 737 - 925 nm, where reflectance was in the order of severity. There were significant differences in reflectance among infestations at wavelengths of 755 and 890 nm particularly. Spectral parameters such as the normalized difference vegetation index (NDVI) and cumulative reflectance may also be used to discriminate levels of infestation. Twelve wavelengths from apparent peaks and valleys of individual spectra were selected as characteristic wavelengths making up the spectral signature of each infestation.     

Microspectrophotometric evidence for two photointerconvertible states of visual pigment in the barnacle lateral eye

Microspectrophotometrically derived difference spectra from the barnacles Balanus amphitrite and B. eburneus show that a blue illumination after an orange illumination causes a decrease in absorption in the blue region and an increase in absorption in the green-yellow region, with an isosbestic point around 535 nm. Orange-following-blue illumination causes the reverse changes. The dark time between the adapting and measuring lights has no influence on the data. The results confirm previously reported ERP measurements which indicate that the barnacle visual pigment has two photointerconvertible dark-stable states. If one assumes a Dartnall nomogram shape for the two absorption spectra, a best fit to the observed difference spectra is obtained with nomograms peaking at 492 nm and 532 nm, with a peak absorbance ratio around 1.6:1. These two nomograms fit very well the ERP action spectra of metarhodopsin and rhodopsin, respectively, thus indicating that the ERP is a reliable measure of visual-pigment changes in the barnacle. The existence of a photostable blue pigment is demonstrated in B. eburneus and in some of B. amphitrite receptors, and the possible influence of this photostable pigment on the various action spectra measured in the barnacle is discussed.     

Excitation spectra of chlorophyll fluorescence in spinach and barley chloroplasts at 4 K

Excitation spectra of chlorophyll a fluorescence in chloroplasts from spinach and barley were measured at 4.2 K. The spectra showed about the same resolution as the corresponding absorption spectra. Excitation spectra for long-wave chlorophyll a emission (738 or 733 nm) indicate that the main absorption maximum of the photosystem (PS) I complex is at 680 nm, with minor bands at longer wavelengths. From the corresponding excitation spectra it was concluded that the emission bands at 686 and 695 nm both originate from the PS II complex. The main absorption bands of this complex were at 676 and 684 nm. The PS I and PS II excitation spectra both showed a contribution by the light-harvesting chlorophyll $\text{a}\text{b}$ protein(s), but direct energy transfer from PS II to PS I was not observed at 4 K. Omission of Mg²⁺ from the suspension favored energy transfer from the light-harvesting protein to PS I. Excitation spectra of a chlorophyll b-less mutant of barley showed an average efficiency of 50–60% for energy transfer from β-carotene to chlorophyll a in the PS I and in the PS II complexes.     

Ceruloplasmin-anion interaction. A circular dichroism spectroscopic study.

The effect of anion binding to ceruloplasmin has been studied using absorption and cirbular dichroism spectral data. At anion to ceruloplasmin molar ratios approaching infinite, OCN-, N3- and SCN- bind to ceruloplasmin giving rise to similar alterations in circular dichroism and absorption spectra. The positive bands at 610 and 520 nm in circular dichroism spectra disappear, a negative one apperars at 600 nm and the peak at 450 nm is only slightly modified. There is a new negative band at 410 nm well-defined in OCN- ceruloplasmin spectra. The decrease in absorption at 610 nm is ascribed to the disruption of one type I Cu-S(cysteine) bond owing presumably to the changes induced by anions in the protein secondary structure. The new band at 410 nm is assigned to a charge transfer transition from the ligand replacing cysteine at its binding site. Both absorption and circular dichroism spectra show isobestic points indicating that anion binding to the enzyme, disruption of one of the two type I Cu-S bonds and coordination of this Cu to another protein residue take place simultaneously.     

Staining method of poly(3-hydroxyalkanoic acids) producing bacteria by nile blue

Summary Using an ethanol solution of nile blue, we have developed an efficient method to detect the colonies of poly(3-hydroxyalkanoic acids) (PHA) producing bacteria on the agar plate. When the bacterial colonies with PHA granules were stained with nile blue, the stained colonies fluoresced bright orange on the irradiation of UV light. In the fluoresce emission spectra, fluorescence intensity increased with an increase in the PHA content of bacterial cells.Alcaligenes eutrophus andA.latus colonies with poly(3-hydroxybutyric acid) (PHB) homopolymer exhibited an emission maximum at 580nm on the excitation at 490nm. On the other hand,Pseudomonas oleovorans andP.putida with medium-chain-length (mcl-) PHA copolymers of C6, C8 and C10 units exhibited an emission maximum at 570nm.     

Some physiological and biochemical changes in marine eukaryotic red tide alga Heterosigma akashiwo during the alleviation from iron limitation

Some physiological and biochemical changes in the marine eukaryotic red tide alga Heterosigma akashiwo (Hada) were investigated during the alleviation from iron limitation. Chlorophyll a/carotenoid ratio increases as a result of iron alleviation. In vivo absorption spectra of iron-limited cells showed a chlorophyll (Chl) absorption peak at 630 nm, 2 nm blue-shifted from the normal position. Low-temperature fluorescence emission spectra of the cells have one prominent Chl emission peak at 685 nm. The cells showed a decrease in fluorescence yield from 685 nm band during alleviation from iron limitation. Low-temperature fluorescence excitation spectra and room-temperature fluorescence spectra indicated an efficient excitation energy transfer in the cells alleviated from iron limitation. Photosynthetic efficiency and carbohydrate content per cell increased after alleviation from iron limitation. Total protein decreased in iron-limited cells, while iron deficiency induced the appearance of specific soluble proteins (17 and 55 kDa).     

The ultraviolet absorption spectra of chromatoid bodies of Entamoeba invadens in situ : An optical titration

Ultraviolet absorption spectra of chromatoid bodies, which have been considered to be crystals of ribosomes, found in the cysts of E. invadens, were obtained by microspectrophotometry. The absorption spectra changed from a single 275 nm peak obtained from fresh cysts to peaks at 260 nm and 310 nm obtained from one day old cysts. Three day old cysts showed a further change to two peaks at 260 nm and 360 nm. We have attributed these changes to the oxidation of tryptophan to formylkynurenine and the hydrolysis of formylkynurenine to kynurenine.     

Red region excitation spectra of protochlorophyllide in dark-grown leaves from plant species with different proportions of its spectral forms

Etiolated leaves of three different species, maize, wheat, and pea, as well as a pea mutant (lip1) were used to compare the excitation spectra of protochlorophyllide (Pchlide) in the red region. The species used have different composition of short-wavelength and long-wavelength Pchlide forms. The relation between different forms was furthermore changed through incubating the leaves in 5-aminolevulinic acid (ALA), which caused an accumulation of short-wavelength Pchlide forms, as shown by changes in absorption and fluorescence spectra. This is the first time a comprehensive comparison is made between excitation spectra from different species covering an emission wavelength range of 675–750 nm using fluorescence equipment with electronic compensation for the variations in excitation irradiance. The different forms of Pchlide having excitations peaks at 628, 632, 637, 650, and 672 nm could be best measured at 675, 700, 710, 725, and 750 nm, respectively. Measuring emission at wavelengths between 675– 710 nm gave an exaggeration of the short-wavelength forms and measuring at longer wavelengths gave for the pea leaves an exaggeration of the 672 nm peak. In general, an energy transfer from short-wavelength Pchlide forms to long-wavelength Pchlide forms occurred, but such an energy transfer sometimes seemed to be limited as a result of a discrete location of the Pchlide spectral forms. The excitation spectra resembling the absorption spectrum most were measured at an emission wavelength of 740 nm. Measuring the excitation at 710 nm gave higher intensity of the spectra but the short-wavelength forms were accentuated.     

Spectral Irradiance and Distribution of Pigments in a Highly Layered Marine Microbial Mat

The spectral irradiance from 400 to 1,100 nm was measured with depth in the intertidal sand mats at Great Sippewissett Salt Marsh, Mass. These mats contained at least four distinct layers, composed of cyanobacteria, purple sulfur bacteria containing bacteriochlorophyll a (Bchl a), purple sulfur bacteria containing Bchl b, and green sulfur bacteria. Spectral irradiance was measured directly by layering sections of mat on a cosine receptor. Irradiance was also approximated by using a calibrated fiber-optic tip. With the tip, irradiance measurements could be obtained at depth intervals less than 250 μm. The irradiance spectra were correlated qualitatively and quantitatively with the distribution of the diverse chlorophyll pigments in this mat and were compared with spectra recorded in plain sand lacking pigmented phototrophs. We found that the shorter wavelengths (400 to 550 nm) were strongly attenuated in the top 2 mm of the mat. The longer wavelengths (red and near infrared) penetrated to much greater depths, where they were attenuated by Bchl a, b, and c-containing anoxygenic phototrophic bacteria. The specific attenuation bands in the irradiance spectra correlated with the specific in vivo absorption bands of the Bchl-protein complexes in the bacteria. We concluded that the pigments in the phototrophs had a profound affect on the light environment within the mat. It seems likely that the diverse Bchl-protein complexes found in the anoxygenic phototrophs evolved in dense mat environments as a result of competition for light.     

Inactivation action spectra of Bacillus subtilis spores with monochromatic soft X rays (0.1-0.6 nm) of synchrotron radiation.

Five types of Bacillus subtilis spores differing in DNA repair and recombinational capacities were exposed in vacuum to monochromatic soft X rays from synchrotron radiation. The inactivation rate constants were obtained from exposure-survival curves upon irradiations at 12 wavelengths in the range of 0.1000 nm (12.40 keV) to 0.6000 nm (2.066 keV). Spores of two repair-deficient strains, UVS (uvrA ssp) and UVP (uvrA ssp polA), exhibited almost equal sensitivities to those of wild-type UVR+, while those of two recombination-deficient strains, RCE (recE) and RCF (recF), exhibited higher sensitivities in the whole wavelength range. This suggested that the repair of DNA damage produced by soft X rays was dependent on the recombinational capabilities. Inactivation action spectra based on photon fluence showed that the effectiveness of the radiation increased as the wavelengths became longer. Abrupt changes in the effectiveness occurred around the wavelengths corresponding to the absorption edges of K-shell electrons of phosphorus and calcium. In both cases, the sensitivity was the highest at the wavelengths of the resonance absorption peak, the next highest at those of the higher energy, and the lowest at the lower energy. Mass energy absorption coefficients of spores were obtained from the transmission of a flake made of spores. They were used to derive inactivation action spectra based on absorbed doses. In these spectra, basal levels of the sensitivity seemed constant, and enhancements of the sensitivity were observed consistent with the absorption by calcium and phosphorus. Thus calcium and phosphorus atoms were the predominant targets for the absorption events leading to the inactivation of spores in the wavelength range examined.     

Biosynthesis of Chlorophyll Regulates Reconstruction of Thylakoid Membrane in Cyanobacterium Synechocystis sp. PCC 6803

By DNA recombination technology in vitro, ORF469- mutant of cynobacterium Synechocystis sp. PCC 6803 was constructed, in which the ORF469 fragment relative to the light-inde-pendent protochlorophyllide (Pchlide) reduction was deleted. In BG-11 medium with 5 mmol/L glucose, the mutant was grown in darkness with a brief period (10 min) of illumination everyday (light-activated heterotrophic growth, LAHG) for 2 weeks to delete chlorophyll (Chl). The 665 mn Chl peak was replaced by the 629 nm Pchlide peak in the absorption spectra of the methanol extracts. The absorption spectra of the intact cells showed only shoulder peak at 620 nm (representing phyco- biliprotein). The thylakoid membrane disappeared, but the amount of phycobilisome did not decrease. When the mutant was transferred from LAHG condition to continuous light illumination for 3 h, the absorbance at 665 nm became higher than that at 629 nm and two peaks at 620 nm and 440 nm,representing phycobiliprotein and Chi-protein complex respectively, appeared in the absorption spectra of the intact cells. Mter exposure to the light for 8 h, the thylakoid membrane was visible in the cells. And for 24 h, a shoulder peak was present at 680 nm in the absorption spectra of the intact cells. Meanwhile the absorption spectra of the methanol extracts had no difference from that of cells grown in the light. Mter 48 h, the shape of the absorption spectra of the intact cells became the same as that of cells grown in the light. The layers of thylakoid membranes were as clear as those of the cells grown in the light. The results indicated that the biosynthesis of chlorophyll regulates the reconstmction of thylakoid membrane rendering the Chl protein complex to play its functional role in photosystems.     

Circular dichroism of rhodopsins and porphyropsins

Circular dichroism and absorption spectra were determined for digitonin extracts of three rhodopsins: cattle, grass frog, and pigeon; and three porphyropsins: channel catfish, bluegill sunfish, and redear sunfish. A comparison of these spectra shows the following: (1) Porphyropsins, like rhodopsins, exhibit two positive CD peaks in the spectral region 321–700 nm: an α peak at about 520 nm and a small β peak at about 355 nm. These peaks substantially diminish upon bleaching. (2) In the CD spectra the α peaks of the porphyropsins are larger than the α peaks of the rhodopsins, while the β peaks are smaller than those of the rhodopsins. This is just the opposite of the corresponding relationship between the peaks in the absorption spectra. (3) The maxima of these peaks in the CD spectra of rhodopsins and porphyropsins are consistently blue-shifted from the corresponding maxima in absorption spectra. (4) Some of the visual pigments show additional positive CD peaks in the spectral region 250–320 nm. In all the visual pigments studied, the CD spectra in this region decrease on bleaching. No reciprocal relationship is observed between any of the CD bands in the visible and near ultraviolet region of the spectrum.     

The effect of eye colour pigments on the action spectrum of Drosophila

In vivo absorption spectra for Drosophila melanogaster eye colour pigment classes (drosopterins and ommatins) were constructed by subtracting the whole eye electroretinographic (ERG) spectral sensitivities of cn and bw respectively from the sensitivities of white-eyed strains. In situ microspectrophotometric (MSP) absorption spectra were also obtained. Both the ERG and MSP drosopterin spectra show a visible peak at 500 nm compared to the 480 nm peak of in vitro drosopterins. For the ommatins, the ERG absorption spectrum peaks at 450 nm while the MSP spectrum peaks at 400 and 525 nm. The ERG spectrum is similar to the in vitro absorption spectrum of xanthommatin while the MSP spectrum is similar to the in vitro absorption spectrum of reduced xanthommatin. The ERG absorption spectra for the drosopterins and the ommatins yield an accurate prediction of the effect of the combined pigments in wild-type eyes. Newly emerged and 7 day post-emergence bw flies show quantitatively similar pigment absorption effects while the drosopterins depress the sensitivity of newly emerged cn flies to a greater extent than that of cn flies 7 days after emergence.     

Biological Synthesis of Semiconductor Zinc Sulfide Nanoparticles by Immobilized Rhodobacter sphaeroides

A novel, clean biological transformation reaction by immobilized Rhodobacter sphaeroides has been developed for the synthesis of zinc sulfide (ZnS) nanoparticles with an average diameter of 8 nm. The nanoparticles were examined by X-ray diffraction, transmission electron microscopy, energy dispersive analyses of X-rays, UV–vis optical absorption and photoluminescence spectra. The average diameter of ZnS nanoparticles varied according to the culture time.