Interactions between photosynthesis and 'light-enhanced dark respiration' (LEDR) in the flagellate Euglena gracilis after irradiation with ultraviolet radiation

The effects of ultraviolet radiation (UV-A, 315-400 nm plus UV-B, 280-315 nm) on photosynthesis and 'light-enhanced dark respiration' (LEDR) in Euglena gracilis have been investigated by using light pulses (80 s) with increasing photon fluence rates of 59, 163, 600, 1180, 2080 and 3340 micromol m(-2) s(-1) and dark periods between the light pulses. LEDR is estimated as the maximum rate of oxygen consumption after a period of light minus the rate of oxygen consumption 30 s after the maximum rate. Without any exposure to UV radiation, the photosynthetic rate and LEDR increase with increasing photon fluence rate. After 20 and 40 min exposures to UV radiation, the photosynthetic rate and LEDR as functions of photon fluence rate are reduced. After a 20 min UV treatment respiration is greater than photosynthesis after the first light pulse of 59 micromol m(-2) s(-1) radiation, and especially at higher photon fluence rates photosynthesis is lower than the control values. The inhibitory effects of UV radiation on photosynthetic rate and LEDR are greater after a 40 min UV exposure than after a 20 min exposure. Only at 600 micromol m(-2) s(-1) is the rate of oxygen evolution greater than that of oxygen consumption after a 40 min UV treatment. Both photosynthetic rate and LEDR are inhibited by the photosynthetic inhibitor DCMU (10(-5) M) in a similar way, which indicates close regulatory interactions between photosynthesis and LEDR. Potassium cyanide (KCN) inhibits dark respiration more than it inhibits LEDR. Dark respiration is not affected to the same degree by UV radiation as are photosynthesis and LEDR.     

PHOTOCHEMISTRY OF THE BISBENZIMIDAZOLE DYE 33258 HOECHST WITH BROMODEOXYURIDINE AND ITS BIOLOGICAL EFFECTS ON BrdUrd-SUBSTITUTED ESCHERICHIA COLI

Abstract— Exposure of BrdUrd-substituted E. coli cells to 360 nm light in the presence of the bisbenzimi-dazole dye 33258 Hoechst increases their sensitivity dramatically. Mutant cells deficient in excision repair of DNA damage ( uur B) are more sensitive than wild type cells, indicating that the cells are able to repair this type of damage. However, they perform only a limited amount of liquid holding recovery (LHR). Exposure of the dye with BrdUrd to near UV light in solution results in the appearance of two BrdUrd derived photoproducts. One appears to be deoxyuridine, and the other — an adduct of BrdUrd-dye. The adduct is acid labile and as a result only uracil is observed in acid-hydrolyzates of DNA after exposure of BrdUrd-substituted cells to 360 nm light in the presence of 33258 Hoechst. The production of uracil is linearly dependent on light exposure. Cells in which 85% of thymidine was replaced by BrdUrd are unable to remove more than 5–10% of uracil from their DNA during postirradiation incubation. However, when only 4% of thymidine is replaced, about 50% of the uracil is removed during 30min incubation after exposure. The results are consistent with our previous work, indicating that BrdUrd interferes with repair via excision-resynthesis. A working hypothesis is suggested to explain this interference.     

PHOTOREACTIVATION OF DNA-CONTAINING CAULIFLOWER MOSAIC VIRUS AND TOBACCO MOSAIC VIRUS RNA ON DATURA

Abstract— Datura stramonium L. is a local lesion host for TMV-RNA and DNA-containing cauliflower mosaic virus (CAMV). Datura can photorepair UV-damaged TMV-RNA and CAMV, giving photoreac-tivation sectors of 0.40 and 0. 33 , respectively. Dose response curves for photoreactivation of TMV-RNA and CAMV show that 4540 min of cool white light (15 W.m^-2) is required for maximum photoreactivation. Blue light and near UV are equally effective in photoreactivating UV-irradiated TMV-RNA, whereas near UV is initially more effective than blue light for the photorepair of UV-inactivated CAMV. Higher doses of near UV apparently inactivate the CAMV photorepair system. In the case of CAMV, photoreactivating light must be applied immediately after inoculation with the virus. Two to three hours of incubation in the dark after inoculation results in complete loss of response to photoreactivating irradiation. In contrast, limited photoreactivation of TMV-RNA occurs even after 4 h of dark incubation after inoculation, although photoreactivating irradiation is most effective when applied immediately after inoculation. Light is required for the maintenance of photoreactivation for both TMV-RNA and CAMV. Daturas placed in the dark for six days lose their ability to photoreacti-vate. Recovery of the TMV-RNA photorepair system is rapid; complete recovery attained with 90 or more min of white light (15 W m-'). Recovery of CAMV photorepair system is slow; 90% recovery attained after only 20 h of light. However, full recovery can be induced by as little as 6h of light when CAMV is inoculated 24 h after the onset of illumination. These results suggest two photorepair systems are present in Datura .     

多钼酸盐–柠檬酸复合膜的多色光致变色

制备了多钼酸盐–柠檬酸光致变色复合膜，紫外光照后发现不同摩尔比的复合膜呈现不同的颜色。当摩尔比为1.0，0.3和0.2时，变色后的薄膜分别显深蓝色，深黄褐色和淡海绿色。通过对薄膜的拉曼光谱分析证实呈现不同的颜色是由于在变色后的膜中生成了不同的物种。柠檬酸在光致变色过程中起着重要的作用, 在紫外光的照射下它作为空穴的捕获剂, 抑制了光生电子和空穴的复合, 使多钼酸盐呈现紫外光致变色现象。     

MULTIPLE EFFECTS OF FLUORESCENT LIGHT ON REPAIR OF ULTRAVIOLET-INDUCED DNA LESIONS IN CULTURED GOLDFISH CELLS

Abstract— It is known that fluorescent light illumination prior to UV irradiation (FL preillumination) of cultured fish cells increases photorepair (PR) ability. In the present study, it was found that FL preillumination also enhanced UV resistance of logarithmically growing cells in the dark. This enhancement of UV resistance differs from induction of PR because it was not suppressed by cycloheximide (CH) and it occurred immediately after FL preillumination. The effects of FL preillumination on repair of UV-induced DNA lesions in the dark were examined by an endonuclease-sensitive site assay to measure the repair of cyclobutyl pyrimidine dimers, and by enzyme-linked immunosorbent assay to quantitate the repair of (6-4) photoproducts. It was found that excision repair ability for (6-4) photoproducts in the genome overall was increased by FL preillumination. Moreover, a decrease in (6-4) photoproducts by FL illumination immediately after UV irradiation of the cells was found, the decrement being enhanced by FL preillumination with or without CH.     

MODE OF COACTION OF PHYTOCHROME AND BLUE LIGHT PHOTORECEPTOR IN CONTROL OF HYPOCOTYL ELONGATION

Abstract The rate of hypocotyl longitudinal growth in seedlings of Sesamum indicum L. is strongly inhibited by continuous blue light (cBL)† and slightly by continuous far-red light while continuous red light (cRL) or red light pulses are hardly effective from 60 h after sowing onwards. Between 36 and 60 h after sowing the growth rate responds to red light pulses the effect of which is fully reversible by long wavelength far-red light. When seedlings are kept in cBL for 3 days and then treated with red light hypocotyl growth rate responds strongly. However, RL effectiveness decreases with time after transfer from BL to RL. BL → darkness transfer experiments with different levels of P_fr established at the beginning of darkness show that after a BL pretreatment phytochrome (P_fr) alone is capable of fully controlling growth rate. When white light (WL) is given no BL effect is detectable in weak WL. Only high light fluxes maintain a typical BL growth rate. At medium WL fluxes elongation rate returns gradually to the dark rate. The simplest explanation of the data is that light absorbed by a separate BL photoreceptor is necessary to maintain responsivity to P_fr. With increasing age of the seedlings the requirement for BL increases strongly. On the other hand, brief light pulses—given to demonstrate photoreversibility of phytochrome—remain equally effective provided that responsivity to P_fr exists.     

Rat lens glycolysis after in vivo exposure to narrow band UV or blue light radiation

UV radiation and short wavelength visible light are known to damage various tissues in the eye. This paper investigates the effect on rat lens glycolysis after in vivo exposure with 90 kJ m⁻² narrow band UV radiation (UVB, 300 nm) and 90 kJ m⁻² blue light (435 nm) radiation. After exposure, all lenses were incubated in Medium 199. Samples of culture medium were withdrawn after 2, 4, 6 h and 5, 10, 20 h in two UVB studies and after 5, 10 and 20 h in a blue light study. Lactate is the major end product of lens glycolysis. Lactate was determined with a modified enzymatic-photometric method. Intralenticular lactate was determined in one UVB experiment. In the UVB experiments we found a lower lactate production in the exposed lenses 2–6 h after exposure. There was an accumulation of lactate inside UVB-exposed lenses after 6 h incubation compared with their contralateral lenses. No significant effect on lactate production was observed in the blue light experiment. Conclusions. UVB induced a reversible inhibition of glycolysis. UVB also induced an accumulation of lactate inside the lens. Blue light tended to increase glycolysis.     

MYCOCHROME SYSTEM AND CONIDIAL DEVELOPMENT IN CERTAIN FUNGI IMPERFECTI

Abstract. A photoreceptor system, “mycochrome”, is involved in a blue and near UV reversible photo-reaction which in turn plays an important role in the photocontrol of conidial development in Alternaria tomato, Botrytis cinerea and Helminthosporium oryzae. Conidial development was controlled by alternating exposures to blue and near UV light with the final response being determined by the final light received. When the final light was near UV a conidium developed; when it was blue, conidiation was inhibited and a “sterile” conidiophore was formed. The effects of the two lights were alternatively reversible. Blue and near UV reversible photoreaction was also found in a light-minus-dark difference spectrum of an intracellular particulate fraction isolated from dark-grown mycelia of A. tomato, and two pigments, P_B (blue absorbing type) and P_NUV (near UV absorbing type), were contained in that photoreaction. P_B was tightly bound to the particulate fraction and P_NUV was either soluble in the cytoplasm or very loosely bound to the particulate fraction. Blue and near UV reversible photoreaction occurred only under the coexistence of P_B and P_NUV. That is, a dip in the near UV region and peak absorbance in the blue region was induced by irradiation with near UV light; however, both conditions were partially dissipated with a subsequent irradiation of blue light. These absorbance changes were revers-ibly repeated by alternating doses of blue and near UV light. When molecular oxygen was introduced after exposure to near UV light, peak absorbance in the blue region rapidly disappeared, and these absorbance changes were repeatedly reversible. Similar results were obtained by treatment with sodium hydrosulfite and molecular oxygen. Thus, it was concluded that the mycochrome system suggested in photo-conidiation was consistent with certain active substances showing blue and near UV reversible absorbance changes and caused by photooxidation reduction reaction.     

PHOTOREACTIVATION and EXCISION REPAIR OF UV INDUCED PYRIMIDINE DIMERS IN THE UNICELLULAR CYANOBACTERIUM GLOEOCAPSA ALPICOLA (SYNECHOCYSTIS PCC 6308)

Abstract— The survival curve obtained after UV irradiation of the unicellular cyanobacterium Synecho-cystis is typical of a DNA repair competent organism. Inhibition of DNA replication, by incubating cells in the dark, increased resistance to the lethal effects of UV at higher fluences. Exposure of irradiated cells to near ultraviolet light(350–500 nm) restored viability to pre-irradiation levels. In order to measure DNA repair activity, techniques have been developed for the chromatographic analysis of pyrimidine dimers in Synechocystis. The specificity of this method was established using a haploid strain of Sacchar-omyces cerevisiae. In accordance with the physiological responses of irradiated cells to photoreactivating light, pyrimidine dimers were not detected after photoreactivation treatment. Incubation of irradiated cells under non-photoreactivating growth conditions for 15 h resulted in complete removal of pyrimidine dimers. It is concluded that Synechocystis contains photoreactivation and excision repair systems for the removal of pyrimidine dimers.     

Adaptation strategies of the sheathed cyanobacterium Lyngbya majuscula to ultraviolet-B

Lyngbya majuscula is a dominant organism in the east coast of India forming characteristic mat in dried saline soils simultaneously exposed to solar radiation of the tropics. Studies on the growth response, changes in the spectral properties of the methanolic extract and protein profile of this estuarine sheathed cyanobacterium to UV-B revealed existence of effective adaptation mechanism to withstand prolonged UV-B radiation. Carotenoids along with MAAs of the organism was increased with increase in UV irradiation. Increase in thickness of the mucilaginous sheath layer as well as cellular carbohydrate content was observed upon exposure to prolonged UV-B dose. Induction of 21 and 33 kDa low molecular weight proteins, and a 99 kDa protein together with formation of distinct multilayered sheath embedding trichomes with granulated cells were the adaptive features of the organism to cope with UV-B stress. The organism was considerably revived after incubating the irradiated cells in mineral medium under florescent light and in the dark suggesting existence of photoreactivation and dark repair in this cyanobacterium. However more experiments are needed to establish the existence of photoreactivation and dark repair mechanism in the studied cyanobacterium.     

Molecular assessment of UVC radiation-induced DNA damage repair in the stromatolitic halophilic archaeon, Halococcus hamelinensis

The halophilic archaeon Halococcus hamelinensis was isolated from living stromatolites in Shark Bay, Western Australia, that are known to be exposed to extreme conditions of salinity, desiccation, and UV radiation. Modern stromatolites are considered analogues of very early life on Earth and thus inhabitants of modern stromatolites, and Hcc. hamelinensis in particular, are excellent candidates to examine responses to high UV radiation. This organism was exposed to high dosages (up to 500 J/m(2)) of standard germicidal UVC (254 nm) radiation and overall responses such as survival, thymine-thymine cyclobutane pyrimidine dimer formation, and DNA repair have been assessed. Results show that Hcc. hamelinensis is able to survive high UVC radiation dosages and that intact cells give an increased level of DNA protection over purified DNA. The organism was screened for the bacterial-like nucleotide excision repair (NER) genes uvrA, uvrB, uvrC, as well as for the photolyase phr2 gene. All four genes were discovered and changes in the expression levels of those genes during repair in either light or dark were investigated by means of quantitative Real-Time (qRT) PCR. The data obtained and presented in this study show that the uvrA, uvrB, and uvrC genes were up-regulated during both repair conditions. The photolyase phr2 was not induced during dark repair, yet showed a 20-fold increase during repair in light conditions. The data presented is the first molecular study of different repair mechanisms in the genus Halococcus following exposure to high UVC radiation levels.     

THE EFFECT OF PROLONGED LIGHT EXPOSURE ON THE EFFECTIVENESS OF PHYTOCHROME IN ANTHOCYANIN SYNTHESIS IN TOMATO SEEDLINGS*

Abstract— The hypocotyl of the tomato ( Lycopersicon esculentum ) seedling synthesizes large amounts of anthocyanin if exposed to prolonged light. Single light pulses are totally ineffective. The involvement of phytochrome can be shown by light pulse treatments following a prolonged light exposure. It is predominantly the action of blue/UV light which leads to a high responsiveness of anthocyanin synthesis towards phytochrome. Moreover, the data suggest a phytochrome-independent action of blue/UV light, in particular of UV-B, on anthocyanin synthesis.     

Photoreactivation in Paramecium tetraurelia under conditions of various degrees of ozone layer depletion

Photoreactivation (PR) is an efficient survival mechanism that helps protect cells against the harmful effects of solar-ultraviolet (UV) radiation. The PR mechanism involves photolyase, just one enzyme, and can repair DNA damage, such as cyclobutane-pyrimidine dimers (CPD) induced by near-UV/blue light, a component of sunlight. Although the balance of near-UV/blue light and far-UV light reaching the Earth's surface could be altered by the atmospheric ozone layer's depletion, experiments simulating this environmental change and its possible effects on life have not yet been performed. To quantify the strength of UVB in sunlight reaching the Earth's surface, we measured the number of CPD generated in plasmid DNA after UVB irradiation or exposure to sunlight. To simulate the increase of solar-UV radiation resulting from the ozone layer depletion, Paramecium tetraurelia was exposed to UVB and/or sunlight in clear summer weather. PR recovery after exposure to sunlight was complete at a low dose rate of 0.2 J/m2 x s, but was less efficient when the dose rate was increased by a factor of 2.5 to 0.5 J/m2 x s. It is suggested that solar-UV radiation would not influence the cell growth of P. tetraurelia for the reason of high PR activity even when the ozone concentration was decreased 30% from the present levels.     

PHOTOREACTIVATION OF ULTRAVIOLET IRRADIATED NON-DIVIDING POPULATIONS OF ICR 2A FROG CELLS

Abstract— Ultraviolet (UV) irradiation of non-dividing populations of ICR 2A frog cells led to their detachment from the surface of the culture dish and eventual lysis. Exposure of the cells to photoreactivating light after UV irradiation prevented cell killing and was accompanied by a loss of endonuclease sensitive sites from DNA. This photoreversal did not take place when the cells were exposed at 4°C to photoreactivating light indicating that the reversal was the result of photoenzymatic repair. As the action of photoreactivating enzyme is specific for the repair of pyrimidine dimers in DNA, these results suggest that pyrimidine dimers in DNA are the critical lesions leading to the death of non-dividing populations of UV irradiated cells.     

Cryptochrome signaling in plants

Cryptochromes are blue light receptors that mediate various light-induced responses in plants and animals. They share sequence similarity to photolyases, flavoproteins that catalyze the repair of UV light-damaged DNA, but do not have photolyase activity. Arabidopsis cryptochromes work together with the red/far-red light receptor phytochromes to regulate various light responses, including the regulation of cell elongation and photoperiodic flowering, and are also found to act together with the blue light receptor phototropins to mediate blue light regulation of stomatal opening. The signaling mechanism of Arabidopsis cryptochromes is mediated through negative regulation of COP1 by direct CRY-COP1 interaction through CRY C-terminal domain. Arabidopsis CRY dimerized through its N-terminal domain and dimerization of CRY is required for light activation of the photoreceptor activity. Recently, significant progresses have been made in our understanding of cryptochrome functions in other dicots such as pea and tomato and lower plants including moss and fern. This review will focus on recent advances in functional and mechanism characterization of cryptochromes in plants. It is not intended to cover every aspect of the field; readers are referred to other review articles for historical perspectives and a more comprehensive understanding of this photoreceptor.     

Impact of the Circadian Clock on UV‐Induced DNA Damage Response and Photocarcinogenesis

The skin is in constant exposure to various external environmental stressors, including solar ultraviolet (UV) radiation. Various wavelengths of UV light are absorbed by the DNA and other molecules in the skin to cause DNA damage and induce oxidative stress. The exposure to excessive ultraviolet (UV) radiation and/or accumulation of damage over time can lead to photocarcinogenesis and photoaging. The nucleotide excision repair (NER) system is the sole mechanism for removing UV photoproduct damage from DNA, and genetic disruption of this repair pathway leads to the photosensitive disorder xeroderma pigmentosum (XP). Interestingly, recent work has shown that NER is controlled by the circadian clock, the body's natural time‐keeping mechanism, through regulation of the rate‐limiting repair factor xeroderma pigmentosum group A (XPA). Studies have shown reduced UV‐induced skin cancer after UV exposure in the evening compared to the morning, which corresponds with times of high and low repair capacities, respectively. However, most studies of the circadian clock–NER connection have utilized murine models, and it is therefore important to translate these findings to humans to improve skin cancer prevention and chronotherapy.     

LOSS OF PHOTOREACTIVATION IN UV-IRRADIATED CULTURED FISH CELLS UNDER DIFFERENT CONDITIONS

Abstract— CAF-MM1 cells derived from a goldfish have photoreactivability for the damage induced by ultraviolet light (UV). When UV-irradiated cells were incubated in the dark at 26_AoC, the longest interval in which photoreactivation (PR) was observed (i.e. effective time for PR), measured by colony formation technique, was about 30 h after the UV irradiation. However, if the cells were incubated at 20_AoC, the effective time was prolonged. Since each time appeared to correspond to the doubling time of the cells at each temperature, the loss of photoreactivability is suggested to be closely related to cell growth or progression of cell cycle. The loss of PR was not observed in the cells held in confluence up to 48 h after UV irradiation, in support of the above suggestion. Photoreactivating enzyme in growing CAF-MM1 cells incubated in the dark for 24 h after UV irradiation was shown to be active, so that it is not possible that the cause of the loss of PR is change in the activity of photoreactivating enzyme.     

SOLVENT-INDUCED PHOTOREVERSIBLE REACTIONS OF C-PHYCOCYANIN FROM SYNECHOCOCCUS SP.

Abstract— C-phycocyanin from Synechococcus sp. ( Anacystis nidulans ) shows photoreversible absorption changes when dissolved in buffer containing 75% ethylene glycol (vol/vol). Irradiation with red light (638 nm) causes a 7.5% decrease in absorbance around the absorption maximum (620 m), while the absorbance around 500 nm increases. Subsequent irradiation with green light (500 nm) partially reverses this change. Final photoreversibility at around 620 nm amounts to ca. 2.5% of the maximum absorbance. These reactions are ascribed to two interconvertible species PC_r and PC_g, the former with a higher absorbance in the red. the latter in the green. The rate of dark reversion from PC_g to PC_r is strongly enhanced by ferricyanide. It is proposed that with this reagent, dark reversion occurs via an oxidized form of PC_g. Furthermore, ferricyanide in the presence of ethylene glycol is capable of reversibly oxidizing part of the chromophores of C-phycocyanin, presumably to a radical. In the absence of ethylene glycol, however, ferricyanide causes total irreversible bleaching of the pigment in the dark. The induced photoreversibility of C-phycocyanin is ascribed to the perturbing action on the protein structure by ethylene glycol in high concentrations. This solvent proved the most suitable perturbant of several compounds tested.     

A NOVEL EFFECT OF UV-B IN A HIGHER PLANT (SORGHUM VULGARE)

Abstract— Two non-photosynthetic photoreceptors (phytochrome and the usual blue/UV light photoreceptor) were previously found to be involved in light-mediated anthocyanin synthesis in the mesocotyl of Sorghum seedlings (Drumm and Mohr, 1978). The decisive point is that phytochrome can act only after a blue/UV light effect has occurred. On the other hand, the expression of the blue/UV light effect is controlled by phytochrome ('obligatory sequential action'). A strong positive interaction between the blue/UV-A and the UV-B part of the spectrum was found, in addition to the above sequential action: an inductive effect of blue or UV-A light can only express itself fully if short wavelength UV (approximately 300–320nm. UV-B range) is also given, either after the blue/UV-A light or simultaneously. Since even small amounts of the UV-B are strongly effective it is probable that this effect plays a role under natural conditions and may not be considered as a mere laboratory artifact.     

UV-induced vertical alignment liquid crystal cell with two species of monomers having vertical-alignment-induced side-chain group and phenanthrene group

We propose the method for formation of the vertical alignment polymer film by polymerisation of the monomers being dissolved in the liquid crystal (LC) material. For obtaining the vertical alignment, combination of the two monomers, 4-(4?-octyloxy-biphenyl-4-yloxy)-butyl ester (AOBBE) and 2,7-dimethacryloyl-oxy-phenanthrene (DMAPhen), was found to be useful.

The monomer DMAPhen initiates the polymerisation under UV light exposure, and the AOBBE unit induces the vertical alignment without generating any alignment defects. The monomer DMAPhen is useful for maintaining the high voltage holding ratio and low residual direct current voltage after UV light exposure because the molecules of DMAPhen do not remain in the LC layer.