Enzymatic and non-enzymatic defense mechanisms against ultraviolet-B radiation in two Anabaena species

Enzymatic and non-enzymatic defense strategies against ultraviolet-B radiation (UV-B, 280–315 nm) were studied in Anabaena doliolum and Anabaena strain L31, two of the most common strains of Indian cyanobacteria. Upon UV-B irradiation, both strains showed a 2–5-fold increase in antioxidative enzymes, such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and peroxidase (POD), as compared to non-irradiated control cultures. These enzymes scavenge damaging reactive oxygen species (ROS), generated by UV-B radiation inside the cells. In addition, these organisms also synthesize mycosporine-like amino acids (MAAs) which are able to carry out UV-screening and/at the same time as UV-quenching. The identification and characterization of three types of MAAs from both Anabaena species were performed using absorption spectroscopy, high performance liquid chromatography (HPLC), electro spray ionization-mass spectrometry (ESI-MS), Fourier Transform Infrared (FTIR) spectroscopy and Nuclear Magnetic Resonance (NMR) spectroscopy. Shinorine was found to be the most common MAA in both Anabaena species while porphyra-334 and mycosporine-glycine were present only in A. doliolum. The results of the present investigation clearly demonstrate that both enzymatic and non-enzymatic defense mechanisms are being employed by A. doliolum and Anabaena strain L31 to counteract the damaging effects of UV-B radiation.     

蓝藻对UV-B增强的响应及其紫外屏蔽物质的研究

紫外线辐射对生物体危害日趋严重,逐渐引起了人们的重视.由于蓝藻在生物进化中的特殊性和在生态系统中的重要性,用于研究UV-B对生物体的影响具有诸多优势,目前国内关于UV-B对蓝藻的影响相关报道较少.本文介绍了近年来国外该领域的相关研究,主要包括UV-B对蓝藻生物量、生理效应,特别是光合作用等方面的影响,同时着重介绍了蓝藻中的紫外吸收物质的研究现状,并进一步探讨了其应用情况.     

Penetration and effects of solar UV-B on phytoplankton and macroalgae

The effects of short wavelength solar radiation on aquatic ecosystems were studied in several marine and freshwater systems. The spectral distribution and the penetration of solar radiation into different water types (coastal and oceanic waters of the Baltic Sea, North Sea, Atlantic and Mediterranean) were investigated. Penetration of short wavelength solar radiation strongly depends on the content of dissolved and particulate substances as well as the concentration of phytoplankton. The primary producers often show a typical vertical distribution within the euphotic zone and are reached as well as affected by the penetrating UV-B radiation. The effect of this radiation was both determined in phytoplankton and macroalgae. Measuring pulse amplitude modulated (PAM) fluorescence indicated that major biomass producers were severely inhibited by surface radiation and even impaired at their natural growth site. Likewise, photosynthetic oxygen production was affected by extended exposure to solar radiation.     

Wavelength-dependent induction of UV-absorbing mycosporine-like amino acids in the red alga Chondrus crispus under natural solar radiation

Polychromatic response spectra for the induction of UV absorbing mycosporine-like amino acids (MAAs) were calculated after exposing small thalli of the red alga Chondrus crispus under various cut-off filters to natural solar radiation on the North Sea island Helgoland, Germany. The laboratory-grown specimens typically contain only traces of palythine and synthesise five different MAAs rapidly and in high concentrations after being transplanted into shallow water. The resulting qualitative and quantitative patterns of MAA induction differed markedly with respect to spectral distribution. Furthermore, the wavebands effective for MAA induction vary within the MAA. UV-B radiation had a negative effect on the accumulation of the major MAAs shinorine (λ_max=334 nm) and palythine (λ_max=320 nm), while short wavelength UV-A exhibits the highest quantum efficiency on their synthesis. In contrast, the synthesis of asterina-330 (λ_max=330 nm), palythinol (λ_max=332 nm) and palythene (λ_max=360 nm) was mainly induced by UV-B radiation. Whether the synthesis of shinorine and palythine is induced by a photoreceptor with an absorption maximum in the short wavelength UV-A and whether a second photoreceptor absorbing UV-B radiation is responsible for the induction of asterina-330, palythinol and palythene remains to be studied.Our results show that C. crispus has a high capacity to adapt flexibly the qualitative and quantitative MAA concentration to the prevailing spectral distribution of irradiance. On one hand, this is regarded as an important aspect with respect to the acclimation of algae to increasing UV-B irradiance in the context of ongoing depletion of stratospheric ozone. On the other hand, the experiment demonstrates that UV-A irradiance is more important for the induction of the major MAAs shinorine and palythine than UV-B.     

Analysis of proteins involved in the production of MAA?s in two Cyanobacteria Synechocystis PCC 6803 and Anabaena cylindrica

Mycosporine- like amino acids (MAAs) are small (<400Da), colourless, water soluble compounds composed of cyclohexenone or cyclohexinimine chromophere conjugated with the nitrogen substituent of amino acid or its amino alcohol. These compounds are known for their UV- absorbing role in various organisms and seem to have evolutionary significance. The biosynthesis of MAAs is presumed to occur via the first part of shikimate pathway. In the present work two cyanobacteria Synechocystis PCC 6803 and Anabaena cylindrica were tested for their ability to synthesize MAAs and protein involved in the production of MAAs. It was found that protein sequence 3-phosphoshikimate 1-carboxyvinyltransferase is involved in producing mycosporine glycine in Synechocystis PCC 6803 and 3-dehydroquinate synthase is involved for producing shinorine in Anabaena cylindrica. Phylogenetic and bioinformatic analysis of Mycosporine like amino acid producing protein sequence of both cyanobacterial species Synechocystis PCC 6803 and Anabaena cylindrica provide a useful framework to understand the relationship of the different forms and how they have evolved from a common ancestor. These products seem to be conserved but the residues are prone to variation which might be due the fact that different cyanobacteria show different physiological process in response of Ultraviolet stress.     

紫外辐射对铜绿微囊藻中类菌孢素氨基酸(MAAs)的诱导效应

用8 w紫外灯,距离藻10 cm,以5 min/d、10 min/d、20 min/d连续照射6 d的间断处理和连续照射1 h、1.5 h、2 h、3h、6 h、12 h的连续处理两种方式处理铜绿微囊藻,测得水溶性色素提取液在200~400 nm间的吸收光谱与对照组相比较,没有产生新的吸收峰,说明MAAs的产生不需要紫外线的诱导。同时随着紫外辐射剂量的增加,MAAs的含量先增加,当达到一定的辐射剂量之后,MAAs的含量随着紫外辐射剂量的增加而减小。     

Ultraviolet and osmotic stresses induce and regulate the synthesis of mycosporines in the cyanobacterium Chlorogloeopsis PCC 6912

The cyanobacterium Chlorogloeopsis PCC 6912 was found to synthesize and accumulate two putative UV sunscreen compounds of the mycosporine (mycosporine-like amino acid; MAA) type: mycosporine-glycine and shinorine. These MAAs were not constitutively present in the cells; their synthesis could be induced specifically either by exposure to UVB radiation (280–320 nm) or by osmotic stress, but not by other stress factors such as heat or cold shock, nutrient limitation, or photooxidative stress. A significant synergistic enhancement of MAA synthesis was observed when both stress factors were applied in combination. Although osmotic stress could induce MAA synthesis, comparison of the intracellular contents of MAAs with those of sugar osmolytes (glucose and trehalose) indicated that MAAs play no significant role in attaining osmotic homeostasis. UVB strongly enhanced the accumulation of shinorine, whereas osmotic stress had a more pronounced effect on mycosporine-glycine. This differential effect on the steady-state contents of each MAA could be explained either by differential regulation of biosynthesis or by differential loss rates of MAAs (leakage) under each condition. A preferential leakage of mycosporine-glycine from the cells after a hypoosmotic shock was detected. The results are interpreted in terms of an adaptive necessity for a combined regulatory control responding to both UV and external osmotic conditions in organisms that accumulate water-soluble sunscreens intracellularly. Received: 26 March 1999 / Accepted: 13 July 1999     

The effects of ultraviolet radiation on photosynthetic performance,growth and sunscreen compounds in aeroterrestrial biofilm algae isolated from building facades

The effects of artificial ultraviolet radiation [UVR; 8 W m⁻² ultraviolet-A (UVA), 0.4 W m⁻² ultraviolet-B (UVB)] on photosynthetic performance, growth and the capability to synthesise mycosporine-like amino acids (MAAs) was investigated in the aeroterrestrial green algae Stichococcus sp. and Chlorella luteoviridis forming biofilms on building facades, and compared with the responses of two green algae, from soil (Myrmecia incisa) and brackish water (Desmodesmus subspicatus). All species exhibited decreasing quantum efficiency (F _v/F _m) after 1–3 days exposure to UVR. After 8–12 days treatment, however, all aeroterrestrial isolates exhibited full recovery under UVA and UVA/B. In contrast, D. subspicatus showed only 80% recovery after treatment with UVB. While Stichococcus sp. and C. luteoviridis exhibited a broad tolerance in growth under all radiation conditions tested, M. incisa showed a significant decrease in growth rate after exposure to UVA and UVA/B. Similarly D. subspicatus grew with a reduced rate under UVA, but UVA/B led to full inhibition. Using HPLC, an UV-absorbing MAA (324 nm-MAA) was identified in Stichococcus sp. and C. luteoviridis. While M. incisa contained a specific 322 nm-MAA, D. subspicatus lacked any trace of such compounds. UV-exposure experiments indicated that all MAA-containing species are capable of synthesizing and accumulating these compounds, thus supporting their function as an UV-sunscreen. All data well explain the conspicuous ecological success of aeroterrestrial green algae in biofilms on facades. Biosynthesis and accumulation of MAAs under UVR seem to result in a reduced UV-sensitivity of growth and photosynthesis, which consequently may enhance survival in the environmentally harsh habitat.     

Biotechnological and industrial significance of cyanobacterial secondary metabolites

Cyanobacteria are considered to be a rich source of novel metabolites of a great importance from a biotechnological and industrial point of view. Some cyanobacterial secondary metabolites (CSMs), exhibit toxic effects on living organisms. A diverse range of these cyanotoxins may have ecological roles as allelochemicals, and could be employed for the commercial development of compounds with applications such as algaecides, herbicides and insecticides. Recently, cyanobacteria have become an attractive source of innovative classes of pharmacologically active compounds showing interesting and exciting biological activities ranging from antibiotics, immunosuppressant, and anticancer, antiviral, antiinflammatory to proteinase-inhibiting agents. A different but not less interesting property of these microorganisms is their capacity of overcoming the toxicity of ultraviolet radiation (UVR) by means of UV-absorbing/screening compounds, such as mycosporine-like amino acids (MAAs) and scytonemin. These last two compounds are true ‘multipurpose’ secondary metabolites and considered to be natural photoprotectants. In this sense, they may be biotechnologically exploited by the cosmetic industry. Overall CSMs are striking targets in biotechnology and biomedical research, because of their potential applications in agriculture, industry, and especially in pharmaceuticals.     

Effects of tropical solar radiation on the motility of filamentous cyanobacteria

Abstract The effects of tropical solar radiation on the motility of the cyanobacteria Anabaena variabilis, Oscillatoria tenuis and two strains of Phormidium uncinatum were studied in Ghana (4.30°N). The percentages of motile filaments were drastically reduced by unfiltered solar radiation. Covering the organisms with various long pass or band pass filters (WG320, GG395 or UG5) revealed that the UV-B, UV-A and visible light components of the solar spectrum were all effective in impairing motility in these organisms. Only partial recovery was observed and only after short exposure times.     

Accessory polypeptides in phycobilisomes of red algae and cyanobacteria

Phycobilisomes of red algae and cyanobacteria contain small amounts of nonpigmented polypeptides in addition to the major constituent biliprotein pigments. The localization of these polypeptides is analyzed by gel electrophoresis of phycobilisome fragments obtained by selective dissociation and subsequent separation. Five groups of biliprotein aggregates are determined, belonging to the 6, 11, 16, 18 and 23 S categories. Accessory nonpigmented high molecular weight proteins (80,000 MW) are exclusively bound to phycobilisome core fractions and thylakoids, thus apparently serving as links between the phycobilisomes and the photosynthetic units of the thylakoids. In contrast, smaller nonpigmented accessory polypeptides of 20,000 to 60,000 MW are preferably found in the peripheral biliprotein stacks. They may either form a compatible link between the phycobilisome core and periphery or bind and co-polymerize with hexameric biliproteins in the peripheral stacks to enhance or effect binding of the aggregates. Furthermore, they may determine the arrangement and composition of the phycobilisomes during development and chromatic adaptation.Abbreviations PE phycoerythrin - PEC phycoerythrocyanin - PC phycocyanin - APC allophycocyanin     

The effect of solar UV-B radiation on terpenes and biomass production in Grindelia chiloensis (Asteraceae), a woody perennial of Patagonia,Argentina

UV-B radiation is absorbed effectively by nucleic acids and other sensitive targets, potentially causing harmful photochemical effects. Protection against UV-B radiation may be afforded by flavonoids and other phenolics, which absorb strongly in the UV region, but little is known about the role played by other compounds, such as terpenes. Grindelia chiloensis, native of Patagonia (Argentina), can accumulate as much as 25% resin (terpenes) in its leaves. The present investigation was carried out to test the effect of solar UV-B radiation on the allocation of photoassimilates to biomass and terpenes. Exposure to UV-B radiation reduced whole plant biomass, plant height and leaf area, and increased leaf thickness and resin accumulation in Grindelia chiloensis. Higher absorbance was found for refined resin in the UV-B waveband from plants grown under solar UV-B radiation than plants without UV-B radiation. These chemical and structural changes could protect the plant from UV radiation, and may help elucidate the importance of epicuticular resins for a species as G. chiloensis native to an environment with maximum daily integrated values of solar UV-B irradiance.     

Electron donors to P700 in cyanobacteria and algae: An instance of unusual genetic variability

Cytochrome c-553 and/or plastocyanin have been isolated from many cyanobacteria and several eukaryotic algae. The isoelectric point for both the cytochrome and plastocyanin varies from that of a basic protein (pI 9.3) in the filamentous cyanobacteria to that of an acidic protein (pI 3.8) in unicellular cyanobacteria and eukaryotes. The cytochrome from a given genus may show isomeric forms distinguishable in either net charge or nonpolar character. Some of the variation in net charge between the cytochromes from different genera is localized in one region of the primary structure.     

Identification of teratogenic polymethoxy-1-alkenes from Cylindrospermopsis raciborskii,and taxonomically diverse freshwater cyanobacteria and green algae

Cylindrospermopsis raciborskii is among the most commonly recognized toxigenic cyanobacteria associated with harmful algal blooms (HAB) in freshwater systems, and specifically associated with multiple water-soluble toxins. Lipophilic metabolites from C. raciborskii, however, were previously shown to exert teratogenicity (i.e. inhibition of vertebrate development) in the zebrafish (Danio rerio) embryo model, specifically suggesting the presence of additional bioactive compounds unrelated to the currently known toxins. In the present study, a series of known teratogenic polymethoxy-1-alkenes (PMA) were identified, purified and chemically characterized from an otherwise well-characterized strain of toxigenic C. raciborskii. Although PMA have been previously identified in other cyanobacteria, this is the first time they have been identified from this recognized HAB species. Following their identification from C. raciborskii, the taxonomic distribution of the PMA was additionally investigated by chemical screening of a freshwater algal (i.e. cyanobacteria, green algal) culture collection. Screening suggests that these compounds are distributed among phylogenetically diverse taxa. Furthermore, parallel screening of the algal culture collection, using the zebrafish embryo model of teratogenicity, the presence of PMA was found to closely correlate with developmental toxicity of these diverse algal isolates. Taken together, the data suggest PMA contribute to the toxicity of C. raciborskii, as well as apparently several other taxonomically disparate cyanobacterial and green algal genera, and may, accordingly, contribute to the toxicity of diverse freshwater HAB.     

Allelopathic effects of Microcystis aeruginosa on green algae and a diatom: Evidence from exudates addition and co-culturing

Allelopathic interactions among phytoplankton species are regarded as one of the important factors contributing to phytoplankton species competition and succession. The role and extent of allelopathic effects of blooming freshwater cyanobacteria on other phytoplankton species in eutrophied waters, however, are still unknown. We examined the allelopathic effect of Microcystis aeruginosa on two common green algae (Scenedesmus quadricauda, Chlorella pyrenoidosa) and a diatom (Cyclotella meneghiniana) by adding exudates from different growth phases and in co-culture tests. Exudates of M. aeruginosa from the exponential growth phase and the stationary phase significantly inhibited the growth of S. quadricauda, C. pyrenoidosa and C. meneghiniana, whereas those from the decline phase increased their growth. The presence of M. aeruginosa extremely inhibited the growth of all tested species in co-cultures within 24 h. Our results indicate that under the tested environmental conditions (25 °C, light 80 μmol quanta m⁻² s⁻¹, manual shaking twice a day), allelopathic effects of M. aeruginosa on other phytoplankton species can significantly contribute to their competitive success.     

Seasonal dynamics of amino acids in two small Siberian reservoirs dominated by prokaryotic and eukaryotic phytoplankton

The comparison of the dynamics of phytoplankton biomass and total amino acid composition was made for two water bodies: in one the phytoplankton were dominated by prokaryotes (i.e., there was a bloom of cyanobacteria) and by eukaryotic microalgae in the other. The dynamics of phytoplankton biomass and of total amino acid composition of water were investigated during the vegetation season. It was found that the only factor that significantly changed the percentages of amino acids in water was the bloom of cyanobacteria in the blooming water body. During the bloom of cyanobacteria, the absolute and relative content of the Leu-Glu group increased, while the contents of other acids generally dropped. Before and after the bloom, no significant variations in the total amino acid composition were recorded. In the reservoir where eukaryotic microalgae dominated, no significant variations in amino acid composition were recorded during the season.     

Strategies of ultraviolet-B protection in microscopic algae

Different species of microalgae show a wide range of susceptibility to ultraviolet-B (UV-B) radiation. To identify factors responsible for the UV-B tolerance of some of the algae, we compared 8 species that are highly tolerant to UV-B to 8 species that are highly susceptible. The tolerant species contained substantial amounts of an acetolysis-resistant residue. The residue consists of sporopollenin, a biopolymer of variable chemical composition that occurs in the algal cell walls and absorbs UV-B radiation. The susceptible species contained little or no sporopollenin. We propose that sporopollenin provides protection to the tolerant species by screening the incident UV-B radiation. Previous studies showed that the mycosporine-like amino acids (MAA) also act as effective UV-B screens. Our data indicate that sporopollenin provides a constant protection while MAA are induced by radiation stress and occur with some delay. The tolerant species also differ from the susceptible species in their capacity to repair the reaction centers damaged by UV-B. The tolerant algae became vulnerable to UV-B when protein synthesis needed for repair was blocked by streptomycin. In the susceptible species, streptomycin had no effect during the UV-B stress. The repair deficiency in the susceptible species can be explained either by relatively less effective protein synthesis or by an inhibition of the protein synthesis by UV-B. In the tolerant species, the structures needed for protein synthesis are protected by UV-B screening of sporopollenin and MAA.     

水生生物的紫外光防护剂--类菌胞素氨基酸

类菌胞素氨基酸（mycosporine—like amino acids,MAAs）是一大类以环己烯酮为基本骨架,与不同类型氨基酸通过缩合作用形成的水溶性物质。它能在真菌、海洋细菌、蓝藻和真核藻类细胞中合成,并在海生无脊椎动物和鱼类等生物体内积累,广泛存在于多种水生生物中。它具有紫外光防护、渗透、繁殖调节及发育保护等功能。本文主要介绍有关类菌胞素氨基酸的分布、结构、生物合成和积累以及生理功能的研究进展。     

Diversity of subaerial algae and cyanobacteria on tree bark in tropical mountain habitats

We report the species composition of subaerial epixylic algae and cyanobacteria from a South-East Asian mountain rainforest locality in Cibodas, West Java. Green algae (Trebouxiophyceae, Chlorophyceae, Trentepohliales) were dominant and Cyanobacteria were the second most frequent group. We specifically concentrated on the comparison of species composition of closed primary forest and open antropogenic spaces. Trentepohliales and Cyanobacteria dominated in open spaces with higher light intensities, whereas closed forest localities were dominated by trebouxiophycean coccal green algae. There was a significantly higher algal diversity in open spaces than in closed forest samples indicating the limiting effect of light on subaerial algal communities of closed tropical forests. A number of isolated strains and morphotypes probably represent undescribed taxa. Presented at the International Symposium Biology and Taxonomy of Green Algae V, Smolenice, June 26–29, 2007, Slovakia.