Phototaxis in the ciliated protozoan Ophryoglena flava: dose-effect curves and action spectrum determination

The sensitivity of positive phototactic orientation of cells of the ciliated protozoan Ophryoglena flava has been measured for white light, broad-band blue and red light, and narrow-band monochromatic light, using a laboratory-developed computer aided system. The white-light fluence rate-response curve shows that there is no negative phototaxis in the fluence rate range investigated (0-15 W/m2) and no adaptation phenomena; it is very well fitted by a hyperbolic function; the fluence rate curves under broad band blue and red light (full width at half maximum, FWHM= 100 nm) can be fitted by the same model. The saturation level is, within experimental errors, the same for the three curves, indicating that there are no chromaticity effects and that if there is more than one photoreceptor pigment, they act independently of each other. The fluence rate-response curves determined under narrow band monochromatic light (FWHM = 10 nm) can also be fitted by the same model and show, within experimental errors, the same saturation level. An action spectrum for positive phototaxis at 10-nm intervals has been calculated from fluence rate-response curves: it shows three maxima, at 420, 540 and 590 nm. This action spectrum is significantly different from the ones for photomotile responses in Blepharisma japonicum, Stentor coeruleus and Chlamydodon mnemosyne, whereas it resembles the ones of Paramecium bursaria and Fabrea salina.     

Evidence for yellow light suppression of lettuce growth

Researchers studying plant growth under different lamp types often attribute differences in growth to a blue light response. Lettuce plants were grown in six blue light treatments comprising five blue light fractions (0, 2, 6% from high-pressure sodium [HPS] lamps and 6, 12, 26% from metal halide [MH] lamps). Lettuce chlorophyll concentration, dry mass, leaf area and specific leaf area under the HPS and MH 6% blue were significantly different, suggesting wavelengths other than blue and red affected plant growth. Results were reproducible in two replicate studies at each of two photosynthetic photon fluxes, 200 and 500 mumol m-2 s-1. We graphed the data against absolute blue light, phytochrome photoequilibrium, phototropic blue, UV, red:far red, blue:red, blue: far red and 'yellow' light fraction. Only the 'yellow' wavelength range (580-600 nm) explained the differences between the two lamp types.     

Photomovement of the Gliding Cyanobacterium Synechocystis sp. PCC 6803

Abstract— Using a computerized videomicroscope motion analysis system, we investigated the photomovements of two Synechocystis sp. (PCC 6803 and ATCC 27184). Synechocystis sp. PCC 6803 displays a relatively slow gliding motion. The phototactic and photokinetic speeds of this cyanobacterium in liquid media were 5μm/min and 15.8 μm/min, respectively, at 3μmol/m²/s of stimulant white light. Synechocystis sp. PCC 6803 senses light direction rather than intensity for phototaxis. Synechocystis sp. ATCC 27184 showed a weak photokinesis but no phototaxis. Analysis of Synechocystis sp. ATCC 27184 suggests that the loss of phototaxis results from spontaneous mutation during several years of subculture. When directional irradiation was applied, the cell population of Synechocystis sp. PCC 6803 began to deviate from random movement and reached maximum orientation at 5 min after the onset of stimulant white light. Synechocystis sp. PCC 6803 showed high sensitivity to the stimulant white light of fluence rates as low as 0.002 |unol/m²/s. Neither 1,3-dichlorophenyldimethyl urea nor cyanide affected phototactic orientation, whereas cyanide inhibited gUding speed. This result suggests that the phototaxis of Synechocystis sp. PCC 6803 is independent of photosynthetic phosphorylation and that its gliding movement is primarily powered by oxidative phosphorylation. In the visible wavelength region, 560 nm, 660 nm and even 760 nm caused positive phototaxis. However, 360 nm light induced strikingly negative phototaxis. Therefore, at least two independent photoreceptors may exist to control phototaxis. The photoreceptor for positive phototaxis appears likely to be a phytochrome-like tetrapyrrole rather than chlorophyll a .     

PHOTOMOVEMENT IN AN "EYELESS" MUTANT OF Chlamydomonas

The function of the stigma ("eyespot") in the green flagellate Chlamydomonas reinhardtii was investigated by comparing the photomovement of the wild-type alga with that of an "eyeless" mutant ( ey 627). Movements of individual cells in response to a blue-green stimulus light were recorded using a videomicroscope system and were analyzed using vectorial methods. Cells of the "eyeless" mutant were phototactic; at a high stimulus fluence rate, their swimming paths were directed away from the light source. Although the orientation of the mutant was not as strongly directional as that of the wild type, it was statistically significant. However, the swimming paths of the mutant cells were very erratic in the presence of the stimulus beam, undergoing frequent changes of direction. Despite the differences in their phototactic orientation, cells of mutant and wild type all showed a distinct step-up photophobic response at the onset of stimulation. These results are consistent with the hypothesis that the stigma plays an accessory role in phototaxis, either by shading the photoreceptor or by acting as a quarter-wave reflector.     

Equal-quantum action spectra indicate fluence-rate-selective action of multiple photoreceptors for photomovement of the thermophilic cyanobacterium Synechococcus elongatus

Unicellular thermophilic cyanobacterium Synechococcus elongatus displayed phototaxis on agar plate at 55 degrees C. Equal-quantum action spectra for phototactic migration were determined at various fluence rates using the Okazaki Large Spectrograph as the light source. The shapes of the action spectra drastically changed depending on the fluence rate of the unilateral monochromatic irradiation: at a low fluence rate (3 mumol/m2/s), only lights in the red region had significant effect; at a medium fluence rate (10 mumol/m2/s), four major action peaks were observed at 530 nm (green), 570 nm (yellow), 640 nm (red) and 680 nm (red). At high fluence rates (30-90 mumol/m2/s), the former two peaks remained, while red peaks at 640 nm and 680 nm disappeared and, interestingly, an action peak around 700-740 nm (far-red) newly appeared. These results indicate that two or more distinct photoreceptors are involved in the phototaxis and that suitable photoreceptors are selectively active in response to the stimulus of light fluence rates. Far-red or red background lights irradiated vertically from above drastically inhibited phototaxis toward red light or far-red light, respectively. These results indicate involvement of some phytochrome(s).     

ACTION SPECTRUM FOR PHOTOREVERSION OF THE ACTIVE Pfr-FRACTION OF Mougeotia in vivo

Abstract— The dichroic oriented fraction of the far-red light absorbing form of phytochrome (Pfr) in the green alga Mougeotia was characterized by action spectroscopy. Microbeam irradiations had to be used for the induction of chloroplast movement in Pfr-containing cells, because of the special dichroic absorption characteristics of the red light absorbing form of phytochrome (Pr) and Pfr in the alga. Fluence-response curves were elaborated especially in the far-red spectral region by reverting Pfr to Pr at the flanks of the cells and thus generating Pfr-gradients. Linearly polarized light vibrating perpendicularly to the cell axis was used, thus corresponding to the S,-transition moments of Pfr at the flanks of the cells. The action spectrum is characterized by a peak at approximately 715 nm and a very pronounced decrease towards 728 and 734 nm. The data indicate that the spectral absorption of the active Pfr-fraction in green Mougeotia is shifted towards shorter wavelengths as compared to extracted phytochrome from etiolated or even green higher plants. This "blue shift" seems to be typical for Pfr from green lower plants.     

Phototaxis in the cyanobacterium Synechocystis sp. PCC 6803: role of different photoreceptors

The second cyanobacterial phytochrome Cph2 from Synechocystis sp. PCC 6803 was suggested as a part of a light-stimulated signal transduction chain inhibiting movement toward blue light. Cph2 has the two bilin binding sites, cysteine-129 and cysteine-1022, that might be involved in sensing of red/far-red and blue light, respectively. Here, we present data on wavelength dependence of the phototaxis inhibition under blue light, indicating that Cph2 itself is the photoreceptor for this blue light response. We found that inhibition of blue-light phototaxis in wild-type cells occurred below the transition point of about 470 nm. Substitution of cysteine-1022 with valine led to photomovement of the cells toward blue light (cph2(-) mutant phenotype). Analysis of mutants lacking cysteine-129 in the N-terminal chromophore binding domain indicated that this domain is also important for Cph2 function or folding of the protein. Furthermore, putative blue-light and phytochrome-like photoreceptors encoded by the Synechocystis sp. PCC 6803 genome were inactivated in wild-type and cph2 knockout mutant background. Our results suggest that none of these potential photoreceptors interfere with Cph2 function, although inactivation of taxD1 as well as slr1694 encoding a BLUF protein led to cells that reversed the direction of movement under blue light illumination in mutant strains of cph2.     

PHOTOTACTIC BEHAVIOR OF INDIVIDUAL CELLS OF CRYPTOMONAS SP. IN RESPONSE TO CONTINUOUS AND INTERMITTENT LIGHT STIMULI

Abstract— The phototactic response of cells of Cryptomonas sp. to stimulation with continuous or intermittent lateral light was determined by an individual cell method using photomicrography and videomicrography. The cells showed positive phototaxis under the conditions studied. The phototactic orientation of individual cells was induced most effectively by irradiation with light of 570 nm; blue light was less effective, and no orientation was found in red light. An intermittent stimulus regime with a long dark interval (250 ms) elicited a weaker phototactic orientation than did a regime with a short dark interval (63 ms) irrespective of the duration of light pulses (16, 250 and 1000 ms). The swimming rate was ca. 240 ums ^-1 and the rotation period ca. 450 ms in the dark, neither of which was greatly affected by stimulation with continuous or intermittent light. Neither step-up nor step-down photophobic responses were observed at the time of onset or removal of the light stimulus under the experimental conditions. The swimming direction of individual cells became gradually oriented toward the light source. Phototactic response was detectable within 4 s after the onset of light stimulation, reaching a saturation level after more than 30 s.     

Action Spectrum for Sporulation and Photoavoidance in the Plasmodium of Physarum polycephalum, as Modified Differentially by Temperature and Starvation

Abstract— The Plasmodium of the myxomycete Physarum polycephalum sporulates in bright natural environments, suggesting a relationship between photobehavior and sporulation. Thus, the action spectra for two light-dependent phenomena as well as the effects of other environmental conditions have been studied. Sporulation like photo-avoidance responded to UVC (near 270 nm) and near IR (near 750 nm) in addition to the well-documented UVA (near 350 nm) and blue (near 460 nm) regions. Sporulation and photoavoidance had similar sensitivities in the shorter wavelengths, while the former was about 100 times more sensitive in near IR. The Plasmodium moved away from light in a wide spectral range. Starvation and high temperature at 31°C (25°C in standard conditions) reduced photoavoidance to UVA and to blue light, respectively. A high fluence rate of UVC suppressed the rhythmic contraction of the Plasmodium, and the action spectrum peaked at 270 nm. These results indicate that the Physarum Plasmodium may stay at brighter places not by positive phototaxis but by weakening the negative phototaxis to sunlight or by other possible taxes such as hydrotaxis. There may be at least four different photo-systems in the Plasmodium.     

Synergistic Effect of Ultrasound and Thiazone–PEG 400 on Human Skin Optical Clearing In Vivo

In this paper, we propose a new physical method in combination with mixed solution of thiazone and polyethylene glycol 400 (thiazone PEG 400 solution) penetration into tissue to assess the skin optical clearing. Four treatments were performed: (1) control group (C); (2) polyethylene glycol 400 (PEG400); (3) 0.25% thiazone (0.25%T); (4) 0.25% thiazone and 5-min ultrasound (0.25%T/SP). The diffuse reflectance spectra and imaging depth of human skin in vivo at different times were measured by spectroscopy and optical coherence tomography (OCT). The optical clearing efficacy of skin was qualitatively and quantitatively analyzed. The results showed that the diffuse reflectance at 540 nm of samples at 10 min after being treated by 0.25%T/SP decreased by approximately 15.51%, whereas, 0.46%, 4.73% and 5.75% were received in C, PEG400 and 0.25%T, respectively. And at 60 min, the decrease in diffuse reflectance of samples in 0.25%T/SP is about 2.22-fold, 1.20-fold compared with that of the samples in PEG 400 and 0.25%T, at 540 nm, respectively. Simultaneously, 0.25%T/SP results in 41.33% increase in OCT 1/e light penetration depth after 60 min. There was a significant difference in the optical clearing effect on skin between ultrasound-mixed solution of thiazone in combination with PEG 400 and the mixed solution (P < 0.05).     

PHOTOBIOLOGIE DU MESOCOTYLE D'AVOINE–IV. SPECTRES D'ACTION ET EFFICACITE QUANTIQUE DE LA LUMIERE

Abstract— The wavelengths most active in influencing the growth of segments of oat mesocotyl are 435 nm (blue), 665 nm (red) and 730 nm (far red). Using a high-energy source (1.5 × 10¹⁸ quanta/cm²), an inhibition in all areas of the spectrum except the far red was obtained. The efficiency of the different radiations was studied: blue light is the most active, red and far red have about the same efficiency; green is a thousand times less active. The effect induced by blue light cannot be directly attributed to phytochrome, because the wavelength of maximum effect does not correspond with that of maximum absorption of the pigment (there is 55 nm difference); also, blue light is more active than the red radiation.     

FLAGELLUM AUTOFLUORESCENCE and PHOTOACCUMULATION IN HETEROKONT ALGAE

Zoids of various brown algae capable of positive or negative phototaxis show strong autofluorescence of their posterior fiagellum and flagellar swelling when excited with blue light of 450 nm. This effect is lacking in zoids without stigma and phototactic capability. Fluorescence studies on living cells point to a flavin as photoreceptor pigment. The spectral sensitivity curve of photoaccumul-ation suggests a carotenoid pigment (maximum 430–450 nm) being involved, with no effect below 380 nm and above 540 nm. The results obtained in this study indicate a flavin as photoreceptor pigment, which is periodically shaded by a carotenoid stigma.     

Color‐Discriminating Retinal Configurations of Sensory Rhodopsin I by Photo‐Irradiation Solid‐State NMR Spectroscopy

SRI (sensory rhodopsin I) can discriminate multiple colors for the attractant and repellent phototaxis. Studies aimed at revealing the color‐dependent mechanism show that SRI is a challenging system not only in photobiology but also in photochemistry. During the photoreaction of SRI, an M‐intermediate (attractant) transforms into a P‐intermediate (repellent) by absorbing blue light. Consequently, SRI then cycles back to the G‐state. The photoreactions were monitored with the ¹³C NMR signals of [20‐¹³C]retnal‐SrSRI using in situ photo‐irradiation solid‐state NMR spectroscopy. The M‐intermediate was trapped at ?40 °C by illumination at 520 nm. It was transformed into the P‐intermediate by subsequent illumination at 365 nm. These results reveal that the G‐state could be directly transformed to the P‐intermediate by illumination at 365 nm. Thus, the stationary trapped M‐ and P‐intermediates are responsible for positive and negative phototaxis, respectively.     

Antagonistic blue- and red-light regulation of cab-gene expression during photosynthetic adaptation in Scenedesmus obliquus.

During adaptation of the photosynthetic apparatus of the green alga Scenedesmus obliquus to various light qualities, the accumulation of chlorophylls and pigment-protein complexes (with specific consideration of chlorophyll a/b-binding (Cab) proteins) and cab-gene expression were determined. The fluence rate dependences for chlorophyll accumulation and cab-gene expression were very different. Very low fluence rates of violet (404 nm), blue (461 nm) and red (650 nm) light below the photosynthetic threshold, i.e. between 10(-3) and 10(-1) mumol m-2 s-1, inhibited all of these reactions in cells grown under heterotrophic conditions. At elevated fluence rates (above 1 mumol m-2 s-1), red light retained its negative regulation, whereas blue light stimulated pigment accumulation. Under autotrophic conditions the pattern was more complex, because chlorophyll accumulation was unaffected by light below the photosynthetic threshold. However, the expression of cab-genes was inhibited by red light but stimulated by blue light. Cells adapted to fluence rates, which ensured photosynthetic energy supply (above 1 mumol m-2 s-1), showed an increase in chlorophyll accumulation, blue light being more effective than red light. The results confirm and extend our previous discovery of two antagonistically acting photoreceptors in Scenedesmus which mediate and coordinate the complex functional and structural changes associated with photosynthetic adaptation. One of these receptor pigments is a blue-light receptor with positive action; the other is a violet-red-light receptor which can operate far below the photosynthetic threshold and exerts a negative regulation.     

BLUE AND GREEN LIGHT EFFECTS,AND THEIR PHOTOREVERSIBILITY,UPON ZOOSPORE PRODUCTION AND THE SYNCHRONIZED CYCLE IN PROTOSIPHON BOTRYOIDES KLEBS*

Abstract —The synchronized asexual cycle in Protosiphon botryoides Klebs is described. A blue light (430 nm) inhibition of zoospore production can be photoreversed by green light (550 nm). Similarly, a green light (550 nm) stimulation of zoospore production can be photoreversed by blue (430 nm) light. 430 nm light also shortens the period of zoospore motility, and 550 nm light lengthens this period.     

Enhanced wavelength-sensitivity of volume holographic grating with dual-photoinitiation system in polymer-dispersed liquid crystal

We propose a method to improve the wavelength-sensitivity of a volume holographic grating by mixing dual-photoinitiation dopants, namely, rose bengal (RB)/N-phenylglycine (NPG) system and methylene blue (MB)/p-toluenesulfonic acid (PTSA) system, which enable the fabrication of gratings upon both green and red light illumination. The RB/NPG green light photoinitiation system is chosen in combination with the red-active MB/PTSA system because RB and MB absorb light near 567 and 665 nm independently. In this case, the holographic gratings, recorded in polymer-dispersed liquid crystal, can be fabricated simultaneously by two different visible laser lights with an output wavelength of 532 and 632.8 nm. The electro-optical performances of the gratings exposed upon 532 and 632.8 nm have been implemented: the diffraction efficiencies (DE) are 75% and 57% respectively when the exposure time is optimised to 2 min and 17 min; the threshold voltage is approximately 2.9 V/μm. The enhanced wavelength-sensitivity of holographic gratings allows for the storage of three-dimensional (3D) images on the same hologram plate, and these 3D images are easily reconstructed by both red and green light.     

BILIRUBIN PHOTOTOXICITY TO HUMAN CELLS BY GREEN LIGHT PHOTOTHERAPY in vitro

Phototherapy of newborn infants with blue or green light is the most common treatment of neonatal hyperbilirubinemia. Using bilirubin bound to human lymphoid and basal skin cells we obtained the green light dose dependency of the bilirubin phototoxicity to these cell types. Cells (3–5× 10⁶/mL) were incubated with bilirubin complexed to human serum albumin (final concentrations 340 μM bilirubin, 150 μM albumin). Under these conditions all cells showed maximum binding of bilirubin. Irradiation with broadband green light (Λ_max= 512 nm) over 24 h led to a light dose-dependent population of cells, which contained no bilirubin on the cell membrane as determined by Nomarski interference microscopy. The light-induced mechanism of the disappearance of bilirubin caused lethal membrane damage to the cells (trypan blue exclusion test). The cell kill rate increased with the irradiation dose and with the fraction of cells with no bilirubin. When 90% of lymphoid cells were bilirubin free, 46% of them were dead (using 480 J cm^?1 green light). Similar results were obtained with basal skin cells. In addition, bilirubin-induced damage of cell membrane and nuclear membrane was also shown by transmission electron microscopy. Bilirubin (340 μM) in the dark led to 5% of the cells being killed. Basal skin cells bind 2.5 times more bilirubin molecules than lymphoid cells and showed a different bilirubin disappearance. Irradiation of bilirubin in carbon tetrachloride with 514.5 nm laser light showed generation of singlet oxygen via its luminescence at 1270 nm. These results demonstrate that green light phototherapy of hyperbilirubinemia may cause both skin and immune system damage.     

Biphasic fluence-response curves and derived action spectra for light-induced absorbance changes in Phycomyces mycelium

The photoresponses of Phycomyces, including phototropism and photocontrol of sporangiophore development, are mediated primarily by blue and UV light. Recent results on these two responses indicated a subsidiary role for green light. We have measured in vivo light-induced absorbance changes (LIAC) in mycelial samples of a caroteneless (carB) strain to compare the effectiveness of UV, blue, and green light. In the dual-wavelength kinetic mode of the spectrophotometer, measuring wavelengths of 445 and 470 nm were chosen, because green light produces substantial absorbance changes between these two wavelengths. Fluence-response curves were measured for 13 wavelengths between 365 and 530 nm, and for variable exposure times between 0.5 and 320 s. With one exception (365 nm), the curves were biphasic. The low fluence component was generally sigmoidal with an abrupt rise. The high fluence component failed to reach saturation for the fluences tested (less than 70 μmol m⁻² s⁻¹). Using the inferred threshold fluences of these two components as criterion effects, we obtained two action spectra. For the low fluence component, the action spectrum showed major peaks at 394, 450, and 530 nm and a minor peak at 416 nm. The high fluence component action spectrum showed very little sensitivity in the blue region. The major sensitivity was in the near UV, and a relatively small peak appeared in the green part of the spectrum at 507 nm. The biphasic character of the fluence-response curves suggests that two photosystems are responsible for the absorbance changes. The low fluence photosystem is sensitive mainly to blue and UV light and may thus represent a physiological blue-light photoreceptor. The high fluence photosystem is clearly not of this type. It (and perhaps the low fluence system as well) may mediate some of the subsidiary physiological effects of green light.     

INHIBITION OF PHOTOTAXIS IN Volvox aureus BY NATURAL AND SIMULATED SOLAR ULTRAVIOLET LIGHT

Exposure to artificial UV wavelengths and the UV component of sunlight delays positive phototaxis in the green alga Volvox aureus. Broad band wavelength filters were used to modify the output from UV-B sources (280-320 nm) and natural sunlight. The delay in phototaxis by artificial UV is increased with exposure to shorter UV-B wavelengths. Natural sunlight experiments were performed with exposure to full sunlight and to its UV component only. The UV component present in summer sunlight produced long periods of inhibition in phototaxis and even lethality, while exposure to the total spectrum of sunlight had no significant effects on movement or survival. The data indicate that although this species of alga is well equipped to deal with present levels of UV exposure, increases in the short UV-B wavelengths in sunlight may force an alteration in patterns of photomovement.     

QUANTUM YIELDS FOR LASER PHOTOCYCLIZATION OF BILIRUBIN IN THE PRESENCE OF HUMAN SERUM ALBUMIN. DEPENDENCE OF QUANTUM YIELD ON EXCITATION WAVELENGTH

The quantum yield for laser photocyclization of bilirubin to lumirubin in the presence of human serum albumin (phi LR) was measured at five monochromatic excitation wavelengths in the range 450-530 nm. Solutions used were optically thin throughout the wavelength range and precautions were taken to exclude contributions from photocyclization of bilirubin XIII alpha impurities. The values obtained (7.2-18 x 10(-4] were lower than those previously reported and showed the following wavelength dependence: 457.9 less than 488.0 less than 501.7 less than 514.5 approximately equal to 528.7. However, the rate of lumirubin formation, normalized to constant fluence, decreased with wavelength over the same wavelength range and no evidence was found that photoisomerization of bilirubin to lumirubin is faster with green (514.5 or 528.7 nm) than with blue (457.9 or 488.0 nm) light. The stereoselectivity of the configurational isomerization of bilirubin to 4Z,15E and 4E,15Z isomers also was studied. This reaction became less regioselective for the 4Z,15E isomer with increasing wavelength. The observed wavelength dependence of phi LR and of the [4E,15Z]: [4Z,15E] ratio at photoequilibrium are consistent with an exciton coupling model in which intramolecular energy transfer can occur between the two pyrromethenone chromophores of the bilirubin molecule in the excited state. Relative rates of lumirubin formation in vivo at different excitation wavelengths and constant fluence were estimated for different optical thicknesses and for different skin thicknesses. These estimates suggest that the recently reported clinical equivalence of blue and green phototherapy lights probably reflects the marked variation of skin transmittance with wavelength more than wavelength-dependent photochemistry. The calculations also indicated that the optimal wavelength for phototherapy is probably on the long wavelength side of the bilirubin absorption maximum.