In vivo nicking and rejoining of nuclear DNA in ultraviolet-irradiated radiation-resistant and sensitive strains of Dictyostelium discoideum

Summary Some aspects of DNA repair in several radiation-resistant and radiation-sensitive strains of Dictyostelium discoideum were investigated by using alkaline sucrose gradients to analyze for the production and resealing of single-strand breaks following irradiation with 254 nm UV. All radiation-resistant strains and all mutants assayed that are sensitive to both UV and ⁶⁰Co gamma rays produced singlestrand breaks in their nuclear DNA after a UV fluence of 15 J/m². Mutants at the radC locus which are sensitive to UV but as resistant as their parental strains to ⁶⁰Co gamma rays produced many fewer single-strand breaks in their DNA after irradiation with UV. Thus, the radC mutations alter a repair pathway specific for UV-induced DNA damage and presumably affect the activity of a UV-damage-specific endonuclease involved in excision repair. All radiation-resistant strains and all of our mutants sensitive to gamma rays rejoined much of their DNA during a three-hour post-UV-irradiation incubation, suggesting that these strains have at least a partially intact excision repair system.Abbreviations used UV ultraviolet light - PBS phosphate buffered saline - cpm counts per minute     

Phosphoinositide-specific phospholipase C forms a complex with 14-3-3 proteins and is involved in expression of UV resistance in fission yeast

The fission yeast plc1 ⁺ gene encodes phosphoinositide-specific phospholipase C. The two- hybrid interaction assay with plexA-plc1 ⁺ as a bait revealed that Plc1p interacted with the 14-3-3 proteins Rad24p and Rad25p. Formation of a complex containing Plc1p and Rad24p in vivo was confirmed by an immunological method. As predicted from the fact that rad24 null mutant cells are hypersensitive to UV irradiation, plc1 null mutant cells were almost as sensitive to UV irradiation as rad24 null mutant cells. In addition, deletion of rad24 in the plc1 null mutant cells did not enhance the UV sensitivity, indicating that plc1 ⁺ and rad24 ⁺ belong to the same epistasis group with respect to UV sensitivity. Whereas Rad24p has been reported to be involved in the DNA damage checkpoint pathway, the delay to mitosis after UV irradiation was not defective either in rad24 null mutant cells or in plc1 null mutant cells in our analysis. Thus, Plc1p is responsible for resistance to UV irradiation, but not for the DNA damage checkpoint pathway, in cooperation with 14-3-3 proteins. Received: 10 July 1997 / Accepted: 15 December 1997     

Reactivation of Giardia lamblia cysts after exposure to polychromatic UV light

Aims: In this study, we determined the ability of a promising alternative UV technology – a polychromatic emission from a medium‐pressure UV (MP UV) technology – to inhibit the reactivation of UV‐irradiated Giardia lamblia cysts. Methods and Results: A UV‐collimated beam apparatus was used to expose shallow suspensions of purified G. lamblia cysts in PBS (pH 7·2) or filtered drinking water to a low dose (1 mJ cm^?2) of MP UV irradiation. After UV irradiation, samples were exposed to two repair conditions (light or dark) and two temperature conditions (25°C or 37°C for 2–4 h). The inactivation of G. lamblia cysts by MP UV was very extensive, and c. 3 log₁₀ inactivation was achieved with a dose of 1 mJ cm^?2. Meanwhile, there was no apparent reactivation (neither in vivo nor in vitro) of UV‐irradiated G. lamblia under the conditions tested. Conclusion: The results of this study indicated that, unlike the traditional low‐pressure (LP) UV technology, an alternative UV technology (MP UV) could inhibit the reactivation of UV‐irradiated G. lamblia cysts even when the cysts were exposed to low UV doses. Significance and Impact of the Study: It appears that alternative UV technology has some advantages over the traditional LP UV technology in drinking water disinfection because of their high level of inactivation against G. lamblia cysts and also effective inhibition of reactivation in UV‐irradiated G. lamblia cysts.     

Inactivation of murine norovirus, feline calicivirus and echovirus 12 as surrogates for human norovirus (NoV) and coliphage (F+) MS2 by ultraviolet light (254 nm) and the effect of cell association on UV inactivation

Aims: To determine inactivation profiles of three human norovirus (NoV) surrogate viruses and coliphage MS2 by ultraviolet (UV) irradiation and the protective effect of cell association on UV inactivation. Methods and Results: The inactivation rate for cell‐free virus or intracellular echovirus 12 was determined by exposure to 254‐nm UV light at fluence up to 100 mJ cm^?2. The infectivity of murine norovirus (MNV), feline calicivirus (FCV) and echovirus 12 was determined by cell culture infectivity in susceptible host cell lines, and MS2 infectivity was plaque assayed on Escherichia coli host cells. The UV fluencies to achieve 4‐log₁₀ inactivation were 25, 29, 30 and 70 (mJ cm^?2) for cell‐free FCV, MNV, echovirus 12 and MS2, respectively. However, a UV fluence of 85 mJ cm^?2 was needed to inactivate intracellular echovirus 12 by 4 log₁₀. Conclusions: Murine norovirus and echoviruses 12 are more conservative surrogates than FCV to predict the UV inactivation response of human NoV. Intracellular echovirus 12 was 2·8‐fold more resistant to UV irradiation than cell‐free one. Significance and Impact of the Study: Variation in UV susceptibilities among NoV surrogate viruses and a likely protective effect of cell association on virus susceptibility to UV irradiation should be considered for effective control of human NoV in water.     

Partial complementation of the UV sensitivity of E. coli and yeast excision repair mutants by the cloned denV gene of bacteriophage T4

Summary The denV gene of bacteriophage T4 was reconstituted from two overlapping DNA fragments cloned in M13 vectors. The coding region of the intact gene was tailored into a series of plasmid vectors containing different promoters suitable for expression of the gene in E. coli and in yeast. Induction of the TAC promoter with IPTG resulted in overexpression of the gene, which was lethal to E. coli. Expression of the TACdenV gene in the absence of IPTG, or the use of the yeast GAL1 or ADH promoters resulted in partial complementation of the UV sensitivity of uvrA, uvrB, uvrC and recA mutants of E. coli and rad1, rad2, rad3, rad4 and rad10 mutants of S. cerevisiae. The extent of denV-mediated reactivation of excision-defective mutants was approximately equal to that of photoreactivation of such strains. Excision proficient E. coli cells transformed with a plasmid containing the denV gene were slightly more resistant to ultraviolet (UV) radiation than control cells without the denV gene. On the other hand, excision proficient yeast cells were slightly more sensitive to killing by UV radiation following transformation with a plasmid containing the denV gene. This effect was more pronounced in yeast mutants of the RAD52 epistasis group.     

The Salmonella typbimurium RecJ function permits growth of P22 abc phage on recBCD+ hosts

Summary We describe recJ mutants of Salmonella typhimurium. The recJ gene maps between sufD and serA (min 62) and is transcribed counterclockwise. Unlike recJ mutants of Escherichia coli, recJ strains of S. typhimurium are sensitive to irradiation with UV light. This sensitivity is equivalent to or greater that that displayed by recBCD mutant strains. The residual ability of phage P22 abc (anti-recBCD) mutants to form plaques on recBCD ⁺ strains is eliminated in recJ hosts. Thus host RecJ function appears to substitute for the anti-RecBCD functions of phage P22 and may serve to limit RecBCD activity.     

Contribution of the UV component of natural sunlight to photoinhibition of photosynthesis in six species of subtidal brown and red seaweeds

Field‐collected specimens of three species of Laminaria and three species of subtidal red algae (Delesseria sanguinea, Plocamium cartilagineum and Phyllophora pseudoceranoides) were exposed to natural summer sunlight on Helgoland (southern North Sea) for up to 4 h at 15 °C. Dark‐adapted variable fluorescence (F_v : F_m) was measured immediately after these treatments, and following 6, 24 and 48 h of recovery in moderate irradiances of white light. The response of plants to the full spectrum of natural sunlight was compared with that to PAR alone, UV‐A + visible, UV‐A + UV‐B, or UV‐A alone. The F_v : F_m values of all species were reduced to minimal values after 4 h in all of these treatments, but those of the more resistant species (Laminaria spp. and P. pseudoceranoides) were higher after shorter exposures to UV radiation alone than to PAR with or without UV. The recovery of F_v : F_m in all species was also more rapid in the two treatments that contained UV radiation alone than in those that included PAR. These results suggest that it is the high irradiances of PAR in natural sunlight which are responsible for the photoinhibition of photosynthesis of subtidal seaweeds and that the current ambient irradiances of UV radiation (either UV‐B or UV‐A) in northern temperate latitudes would not contribute significantly to this photoinhibition.     

The RAD3 gene of Saccharomyces cerevisiae: isolation and characterization of a temperature-sensitive mutant in the essential function and of extragenic suppressors of this mutant

Summary Mutations in the RAD3 gene of Saccharomyces cerevisiae were generated by integration of a mutagenized incomplete copy of the cloned gene into wild-type cells. Integrants were mass screened for colonies with abnormal growth characteristics at 37°C. A single temperature-sensitive mutant (rad3ts-1) was isolated and was shown to result from a missense mutation at codon 73 of the RAD3 gene. When shifted from 30° C to 37° C the strain undergoes only 2–4 cell doublings. This phenotype can be rescued by plasmids in which the essential function of the cloned RAD3 gene is intact, but not plasmids in which this function is inactivated. The mutant strain is weakly sensitive to ultraviolet (UV) radiation at restrictive temperatures. Measurement of RNA, DNA and protein synthesis at various times after shifting to restrictive temperatures does not show preferential inactivation of any one of these parameters and the temperature-sensitive mutation does not cause arrest at any specific phase of the cell cycle. The rad3ts-1 strain was transformed with multicopy plasmids from a normal yeast genomic library and two plasmids that partially suppress the temperature-sensitive phenotype were isolated. These suppressor genes (designated SRE1 and SRE2) are distinct from RAD3 and do not suppress the phenotype of several other temperature-sensitive mutants tested. Mutant strains carrying disruptions of the SRE1 gene are viable and are not sensitive to UV or radiation.     

Bacterial endospores and their significance in stress resistance

In terms of resistance to extreme environmental stresses, the bacterial spore represents a pinnacle of evolution. Spores are highly resistant to a wide variety of physical stresses such as: wet and dry heat, UV and gamma radiation, oxidizing agents, chemicals, and extremes of both vacuum and ultrahigh hydrostatic pressure. Some of the molecular mechanisms underlying spore resistance properties have been elucidated in the laboratory, and involve both: (i) protection of vital spore macromolecules during dormancy, and (ii) repair of damaged macromolecules during germination. Our group has recently become interested in testing if the laboratory model of spore UV resistance is relevant to spore persistence in the environment. We have constructed a number of Bacillus subtilis strains which are defective in various DNA repair systems and spore structural components. Using spores of these strains, we have been exploring: (i) the types of damage induced in DNA by the UV-B and UV-A components of sunlight; (ii) the relative contribution of the major spore DNA repair systems to spore solar radiation resistance; and (iii) the role of spore structural components such as the spore coats and dipicolinic acid (DPA) in attenuation of the lethal and mutagenic effects of solar UV. The current data are reviewed with the ultimate goal of obtaining a complete model describing spore persistence and longevity in the terrestrial solar UV radiation environment. This revised version was published online in August 2006 with corrections to the Cover Date.     

Functional analysis of theDrosophila CDC2 Dm gene in fission yeast

Thecdc2 ⁺ gene product (p34^cdc2) is a protein kinase that regulates entry into mitosis in all eukaryotic cells. The role that p34^cdc2 plays in the cell cycle has been extensively investigated in a number of organisms, including the fission yeastSchizosaccharomyces pombe. To study the degree of functional conservation among evolutionarily distant p34^cdc2 proteins, we have constructed aS. pombe strain in which the yeastcdc2 ⁺ gene has been replaced by itsDrosophila homologue CDC2Dm (theCDC2Dm strain). ThisCDC2Dm S. pombe strain is viable, capable of mating and producing four viable meiotic products, indicating that the fly p34^CDC2Dm recognizes all the essentialS. pombe cdc2 ⁺ substrates, and that it is recognized by cyclin partners and other elements required for its activity. The p34^CDC2Dm protein yields a lethal phenotype in combination with the mutant B-type cyclin p56^cdc13-117, suggesting that thisS. pombe cyclin might interact less efficiently with theDrosophila protein than with its native p34^cdc2 counterpart. ThisCDC2Dm strain also responds to nutritional starvation and to incomplete DNA synthesis, indicating that proteins involved in these signal transduction pathways, interact properly with p34^CDC2Dm (and/or that p34^cdc2-independent pathways are used). TheCDC2Dm gene produces a ‘wee’ phenotype, and it is largely insensitive to the action of theS. pombe weel ⁺ mitotic inhibitor, suggesting thatDrosophila weel ⁺ homologue might not be functionally conserved. ThisCDC2Dm strain is hypersensitive to UV irradiation, to the same degree asweel-deficient mutants. A strain which co-expresses theDrosophila and yeastcdc2⁺ genes shows a dominantwee phenotype, but displays a wild-type sensitivity to UV irradiation, suggesting that p34^cdc2 triggers mitosis and influences the UV sensitivity by independent mechanisms. Communicated by B. J. Kilbey     

Loss of heterozygosity is induced in <Emphasis Type="Italic">Candida albicans</Emphasis> by ultraviolet irradiation

Candida albicans is a human fungal pathogen and has been extensively studied because of its clinical importance. Comprehensive gene analyses have, however, made little progress. This is because of the diploid and asexual characteristics of the fungus that hamper gene disruptions. In this study, we found that ultraviolet (UV) irradiation, as well as mutagen treatment, strongly stimulated loss of heterozygosity (LOH) in strains harboring artificially constructed heterozygosity. UV-induced LOH occurred more frequently in cells within the logarithmic phase of growth compared to those within the stationary phase of growth. This was observed at all loci tested on chromosome 7, except for a locus neighboring the centromere. C. albicans RAD52, whose orthologue in Saccharomyces cerevisiae was reported to be involved in DNA repair by homologous recombination, was shown to be required for UV-induced LOH. These results suggest that high efficiency LOH caused by UV irradiation could be a prominent tool for gene analyses in C. albicans.     

Diversity in UV sensitivity and recovery potential among bacterioneuston and bacterioplankton isolates

Aims: To assess the variability in UV‐B (280–320 nm) sensitivity of selected bacterial isolates from the surface microlayer and underlying water of the Ria de Aveiro (Portugal) estuary and their ability to recover from previous UV‐induced stress. Methods and Results: Bacterial suspensions were exposed to UV‐B radiation (3·3 W m^?2). Effects on culturability and activity were assessed from colony counts and ³H‐leucine incorporation rates, respectively. Among the tested isolates, wide variability in UV‐B‐induced inhibition of culturability (37·4–99·3%) and activity (36·0–98·0%) was observed. Incubation of UV‐B‐irradiated suspensions under reactivating regimes (UV‐A, 3·65 W m^?2; photosynthetic active radiation, 40 W m^?2; dark) also revealed diversity in the extent of recovery from UV‐B stress. Trends of enhanced resistance of culturability (up to 15·0%) and enhanced recovery in activity (up to 52·0%) were observed in bacterioneuston isolates. Conclusions: Bacterioneuston isolates were less sensitive and recovered more rapidly from UV‐B stress than bacterioplankton isolates, showing enhanced reduction in their metabolism during the irradiation period and decreased culturability during the recovery process compared to bacterioplankton. Significance and Impact of the Study: UV exposure can affect the diversity and activity of microbial communities by selecting UV‐resistant strains and alter their metabolic activity towards protective strategies.     

Effect of ultraviolet radiation on cell division and microtubule organization inPetunia hybrida protoplasts

Summary Mesophyll protoplasts isolated fromPetunia hybrida were subjected to UV radiation (280–360 nm) in an attempt to assess whether (a) UV radiation has an effect on cortical microtubule organization, (b) UV radiation affects the progression of protoplasts through the cell cycle, and (c) there is a connection between the effect of UV radiation on cell division and the polymerization state of the microtubules. The proto plasts were irradiated with the following UV doses: 4, 8, 12, and 24mmol photons/m², 30 min after isolation. Cell cycle analysis and immuno-localization of microtubules were carried out 0, 24, 48, and 72 h after irradiation. The length of cortical microtubules was determined after irradiation and in corresponding controls. We found that UV radiation induced breaks in cortical microtubules resulting in shorter fragments with increasing dose. Also, the protoplasts were delayed in their progression through the cell cycle, with G1 and G2 phases being affected as well as the S phase. The commencement of DNA synthesis in the irradiated protoplasts followed the re-establishment of a microtubule network. At 48 h after irradiation the protoplasts in all treatments, except for the 24 mmol/m², had cortical microtubules of similar length, and at 72 h after irradiation only the protoplasts that had received 24 mmol photons/m² had not started dividing.Abbreviations BSA bovine serum albumin - DMSO dimethyl sulfoxide - FDA fluorescein diacetate - MT microtubules - MTSB microtubule stabilizing buffer - PAR photosynthetically active radiation (400–700 nm) - PBS phosphate buffered saline - UV ultraviolet     

产毒与不产毒铜绿微囊藻对模拟酸雨及紫外辐射的生理响应

为了比较研究酸雨与紫外辐射对淡水水体常见藻华蓝藻的生理学影响,选取铜绿微囊藻(Microcystis aeruginosa)产毒(FACHB-905)与不产毒(FACHB-469)株系作为实验材料,通过人工模拟酸雨,研究了不同p H处理后2藻株的光合生理变化以及对紫外辐射的敏感性的异同。实验设置3个p H梯度,p H7.10为对照组(正常培养基培养的藻体),两模拟酸雨处理组(p H5.65和p H4.50);两种辐射处理,可见光处理(PAR)以及全波长辐射处理(PAB)。研究结果表明,905藻株细胞粒径在各p H处理下都要显著高于469藻株,模拟酸雨处理显著降低了两藻株细胞的平均粒径及体积,但叶绿素含量显著提高;酸雨处理同时也引起细胞死亡率的增加,表现为藻体有效光化学效率显著降低,生长速率显著受到抑制,低p H下呈负增长,且这种抑制程度在469下更为显著。高的可见光以及紫外辐射处理,使两株系有效光化学效率随p H的降低而呈降低趋势,其中469藻株降低至更低的水平,且高光辐射以及紫外诱导的抑制率要显著高于905藻体,这可能与469藻株较低的光保护色素有关(较低的类胡萝卜素以及紫外吸收物质)。在未来全球变化背景下,不同种类的浮游植物对环境变化的响应及适应能力不同,可改变水体的群落结构和种群丰度,铜绿微囊藻905较469较强的耐受酸雨以及紫外辐射的能力,可能会使该株系在竞争力上占据优势。     

The genetic conditions in heterozygous and homozygous populations of Drosophila. II. X-ray induced lethals in a homozygous and a heterozygous population

Strains of Drosophila melanogaster were made isogenic for their second chromosomes by means of the marker strain LCy/Pm. One of these strains was used as a founder for a homozygous experimental population (W). All other strains were mixed and established a heterozygous population (LKW). Both populations were free of lethals in the beginning with respect to their second chromosomes. After they had been exposed to an X-ray irradiation of 7000 r they contained about 26 per cent newly induced lethal chromosomes. Whereas in the heterozygous population the lethal frequency decreased rather fast to 10 per cent, that of the homozygous population remained rather constant at 25 per cent during a period of 135 days. After a year of continuation, however, both populations reached the same lethal frequency of about 10 per cent. Allelism tests carried out after 10 generations revealed that there was a highly heterotic lethal factor in the homozygous population. After excluding this heterotic lethal from the calculations, the lethal frequencies of the two populations remained significantly different. It was assumed that the relative mean fitness of lethal heterozygotes was generally higher in the homo-than in the heterozygous populations. The results indicate that homozygous populations are much more capable of incorporation new mutations than heterozygous.     

Developmental changes in the sensitivity of Volvox to ultraviolet light

The response of Volvox to ultraviolet irradiation was analyzed. Young individuals isolated from a synchronous culture were exposed to UV light (120 J/m²) and subjected to variable lenght periods of dark following irradiation. The major effect of the UV treatment was the inability of the gonidia present in the colonies at the time of irradiation to continue and complete the developmental program. Individuals show a heightened sensitivity to UV for a limited period immediately following inversion and are insensitive at other stages of development. The cytotoxic effect of UV during this interval is completely reversed by the immediate exposure to white light and is increased with longer periods of dark treatment prior to exposure to white light. The temporal profile of the sensitivity defines a smooth curve in which the maximal sensitivity occurs three hours after inversion. The response to higher doses of UV (up to 500 J/m²) is a nonlinear increase in cytotoxicity and is disproportionanately greater in those individuals just prior to the period of maximal sensitivity than those later in development. The results suggest that Volvox has at least two pathways for the repair of UV damage and that one of these, the principal dark repair pathway, is temporarily deficient in the gonidia of young individuals.     

APOPTOTIC‐LIKE CELL DEATH PATHWAY IS INDUCED IN UNICELLULAR CHLOROPHYTE CHLAMYDOMONAS REINHARDTII (CHLOROPHYCEAE) CELLS FOLLOWING UV IRRADIATION: DETECTION AND FUNCTIONAL ANALYSES1

Chlamydomonas reinhardtii (Ehrenberg) cells exhibited cell death process akin to that of apoptosis when exposed to ultraviolet (UV)‐C irradiation (1–100 J/m²). We observed typical hallmarks of apoptosis including cell shrinkage, associated nuclear morphological changes, flipping of phosphatidylserine, and DNA fragmentation detected by the terminal deoxynucleotidyl transferase‐mediated dUTP nick end‐labeling assay and oligonucleosomal DNA laddering assay. Interestingly, fluorescence imaging of DNA changes in UV‐C exposed cells, following PicoGreen staining, revealed that extra‐nuclear DNA disintegrates before that of nuclear changes, where the latter extensively diffuses out of the nuclear compartment, spreading into the whole cell and reaching the periphery of dying cells. Antibodies against a mammalian caspase‐3 shared epitopes with a protein of 28 kDa; whose pattern of expression correlated with the onset of cell death. Moreover, growth experiments indicate that spent medium recovered from UV‐C exposed cells exhibit a protective effect against cell killing of fresh cultures of C. reinhardtii cells by UV irradiation. The protective effect of UV‐spent medium is not a general growth promotional response on normal cells, but rather, is specific to UV‐exposed cells. We propose a model that C. reinhardtii cells exposed to UV elicit apoptotic‐like changes, which in turn lead to an adaptive response in neighboring cells against fresh rounds of UV exposure, thereby promoting survival of the cell population.     

Combination of high‐frequency ultrasound and UV radiation of excilamp for surface disinfection

The combination of high‐frequency ultrasound (HFUS) and UV represents a new approach to disinfecting surfaces. This study aimed to examine the inactivation efficiency of HFUS (1.7 MHz) and monochromatic UV radiation of KrCl excilamp (222 nm) in a single and a sequential mode against Bacillus cereus cells and spores added to glass surfaces. When treated by UV only, cells at populations of 10³, 10⁴, and 10⁵ colony‐forming units (CFU)/cm² showed 100% disinfection at high doses up to 1760 mJ/cm². Spores at 10⁴ CFU/cm² were completely inactivated at a dose of 1170 mJ/cm². Treatment with aqueous aerosol (produced by HFUS) reduced cell counts by 100% within a 40‐min exposure, whereas it was ineffective in inactivating spores under these conditions. In a sequential mode, the contaminated surface was pretreated with the sonicated aqueous aerosol and subsequently irradiated with the excilamp. It was found that HFUS exposure times and UV doses for complete inactivation decreased by a factor of 2 and 6–7, respectively, compared to sole HFUS or UV. A portable apparatus for surface disinfection was designed. The combined HFUS/UV method may be a promising technique for rapid disinfection of microbially contaminated surfaces.     

pEffects of UV radiation on the ultrastructure of several red algae

The effect of ultraviolet (UV) radiation on the ultrastructure of four red algae, the endemic Antarctic Palmaria decipiens (Reinsch) Ricker and Phycodrys austrogeorgica Skottsberg, the Arctic‐cold temperate Palmaria palmata (Linnaeus) O. Kuntze and the cosmopolitan Bangia atropurpurea (Roth) C. Agardh was studied. All four species showed a formation of ‘inside‐out’ vesicles from the chloroplast thylakoids upon exposure to artificial UV‐radiation. In P. decipiens, most vesicles were developed after 8 h and in P. palmata, after 48 h of UV exposure. In B. atropurpurea, vesi‐culation of thylakoids was observed after 72 h of UV irradiation. In Ph. austrogeorgica, the chloroplast envelope and thylakoid membranes were damaged and the phycobilisomes became detached from the thylakoids after 12 h of UV exposure. Ultraviolet‐induced changes in the membrane structure of mitochondria were observed in P. decipiens and P. palmata. However, in P. decipiens they were reversible as was the damage in chloroplast fine structure after 12 h of UV treatment. Protein crystals in Ph. austrogeorgica showed degradation after exposure to UV radiation. Different methods of fixation and embedding macroalgal material are discussed. These findings give insight into the fine structural changes which occur during and after UV exposure and indicate a relationship between the species dependent sensitivity to UV‐exposure and the depth distribution of the different species.     

The interaction of visible and UV-B light during photodamage and repair of Photosystem II

In order to understand the mechanism of photodamage induced by solar radiation under natural conditions, we studied the interaction of visible and ultraviolet-B light in the inactivation and repair of the Photosystem II complex by using oxygen evolution and flash-induced chlorophyll fluorescence measurements. In isolated spinach thylakoids and Synechocystis 6803 cells, in which de novo protein synthesis is blocked by lincomycin, photodamage of Photosystem II by visible and UV-B light is characterized by linear semilogarithmic inactivation curves for both separate and combined illumination protocols. The extent of PS II inactivation obtained after combined illumination can be well simulated by assuming independent damaging events induced by visible and UV-B photons. In intact Synechocystis cells capable of protein repair, simultaneous illumination by visible and UV-B light impairs Photosystem II activity to a smaller extent than expected from the independent damaging events. This protective effect is pronounced at low visible light (130 μE m⁻² s⁻¹), but becomes negligible at high intensities (1300 μE m⁻² s⁻¹). Exposure of intact Synechocystis 6803 cells to direct sunlight leads to a rapid inactivation of PS II, accompanied by the accumulation of donor side inhibited centers. This phenomenon, which shows the impairment of the manganese cluster of water oxidation was not observed when the ultraviolet components of sunlight were filtered out. We conclude that visible and UV-B photons inactivate PS II via non-interacting mechanisms, which affect different target sites. In intact cells, the two spectral regions do interact, and results in synergistically enhanced protein repair capacity when UV-B radiation is accompanied by low intensity visible light, which provides protection against photodamage. However, this ameliorating effect becomes insignificant at high light intensities characteristic of direct sunlight. This revised version was published online in June 2006 with corrections to the Cover Date.