The sensitivity to UV‐B radiation (UVBR: 280–315 nm) was tested for littoral (Palmaria palmata[L.] O. Kuntze, Chondrus crispus Stackhouse) and sublittoral (Phyllophora pseudoceranoides S. G. Gmelin, Rhodymenia pseudopalmata[Lamouroux] Silva, Phycodrys rubens[L.] Batt, Polyneura hilliae[Greville] Kylin) red macrophytes from Brittany, France. Algal fragments were subjected to daily repeated exposures of artificial UVBR that were realistic for springtime solar UVBR at the water surface in Brittany. Growth, DNA damage, photoinhibition, and UV‐absorbing compounds were monitored during 2 weeks of PAR + UV‐A radiation (UVAR) + UVBR, whereas PAR + UVAR and PAR treatments were used as controls. The littoral species showed a higher UV tolerance than the sublittoral species. After 2 weeks, growth of P. palmata and C. crispus was not significantly affected by UVBR, and DNA damage, measured as the number of cyclobutane‐pyrimidine dimers per 106 nucleotides, was negligible. Photoinhibition, determined as the decline in optimal quantum yield, was low and decreased during the course of the experiment, coinciding with the production of UV‐absorbing compounds in these species. In contrast, no UV‐absorbing compounds were induced in the sublittoral species. Growth rates of P. pseudoceranoides and R. pseudopalmata were reduced by 40% compared with the PAR treatment. Additionally, constant levels of DNA damage and pronounced photoinhibition were observed after the UVBR treatments. Growth was completely halted for Phycodrys rubens and Polyneura hilliae, whereas DNA damage accumulated in the course of the experiment. Because Phycodrys rubens and Polyneura hilliae showed the same degree of photoinhibition as the other sublittoral species, it appears that the accumulation of DNA damage may have been responsible for the complete inhibition of growth. The results suggest an important role of DNA repair pathways in determining the UV sensitivity in red macrophytes.
Abstract. The hydrographic properties of the KongsfjordenKrossfjorden system (79 • N, Spitsbergen) are affected by Atlantic water incursions as well as glacier meltwater runoff. This results in strong physical gradients (temperature, salinity and irradiance) within the fjords. Here, we tested the hypothesis that glaciers affect phytoplankton dynamics as early as the productive spring bloom period. During two campaigns in 2007 (late spring) and 2008 (early spring) we studied hydrographic characteristics and phytoplankton variability along two transects in both fjords, using highperformance liquid chromatography (HPLC)-CHEMTAX pigment fingerprinting, molecular fingerprinting (denaturing gradient gel electrophoresis, or DGGE) and sequencing of 18S rRNA genes. The sheltered inner fjord locations remained colder during spring as opposed to the outer locations. Vertical light attenuation coefficients increased from early spring onwards, at all locations, but in particular at the inner locations. In late spring meltwater input caused stratification of surface waters in both fjords. The inner fjord locations were characterized by overall lower phytoplankton biomass. Furthermore HPLC-CHEMTAX data revealed that diatoms and Phaeocystis sp. were replaced by small nano-and picophytoplankton during late spring, coinciding with low nutrient availability. The innermost stations showed higher relative abundances of nano-and picophytoplankton throughout, notably of cyanophytes and cryptophytes. Molecular fingerprinting revealed a high similarity between inner fjord samples from early spring and late spring samples from all locations, while outer samples from early spring clustered separately. We conclude that glacier influence, mediated by early meltwater input, modifies phytoplankton biomass and composition already during the spring bloom period, in favor of low biomass and small cell size communities. This may affect higher trophic levels especially when regional warming further increases the period and volume of meltwater.
The response of the bipolar moss Sanionia uncinata (Hedw.) Loeske to ambient and enhanced UV-B radiation was investigated at an Antarctic (Léonie Island, 67 °°°° 35 ′ ′ ′ ′ S, 68 °°°° 20 ′ ′ ′ ′ W) and an Arctic (Ny-Alesund, 78 °°°° 55 ′ ′ ′ ′ N, 11 °°°° 56 ′ ′ ′ ′ E) site, which differed in ambient UV-B radiation (UV-BR: 280-320 nm) levels. The UV-BR effects on DNA damage and photosynthesis were investigated in two types of outdoor experiments. First of all, sections of turf of S. uncinata were collected in an Arctic and Antarctic field site and exposed outdoors to ambient and enhanced UV-BR for 2 d using UV-B Mini-lamps. During these experiments, chlorophyll a fluorescence, chlorophyll concentration and cyclobutyl pyrimidine dimer (CPD) formation were measured. Secondly, at the Antarctic site, a long-term filter experiment was conducted to study the effect of ambient UV-BR on growth and biomass production. Additionally, sections of moss turf collected at both the Antarctic and the Arctic site were exposed to UV-BR in a growth chamber to study induction and repair of CPDs under controlled conditions. At the Antarctic site, a summer midday maximum of 2·1 W m − − − − 2 of UV-BR did not significantly affect effective quantum yield ( ∆ ∆ ∆ ∆ F / F m ′ ′ ′ ′ ) and the ratio of variable to maximal fluorescence ( F v / F m ). The same was found for samples of S. uncinata exposed at the Arctic site, where summer midday maxima of UV-BR were about 50% lower than at the Antarctic site. Exposure to natural UV-BR in summer did not increase CPD values significantly at both sites. Although the photosynthetic activity remained largely unaffected by UV-B enhancement, DNA damage clearly increased as a result of UV-B enhancement at both sites. However, DNA damage induced during the day by UV-B enhancement was repaired overnight at both sites. Results from the long-term filter experiment at the Antarctic site indicated that branching of S. uncinata was reduced by reduction of ambient summer levels of UV-BR, whereas biomass production was not affected. Exposure of specimens collected from both sites to UV-BR in a growth chamber indicated that Antarctic and Arctic S. uncinata did not differ in UV-BR-induced DNA damage. It was concluded that S. uncinata from both the Antarctic and the Arctic site is well adapted to ambient levels of UV-BR.
We studied the physiological response of phytoplankton to the interacting effects of 3 factors affected by global climate change: CO 2 , nutrient loading and irradiance. Treatments had a high and low level for each factor: CO 2 was bubbled at 1000 ppm by volume versus present atmo spheric values; high nutrient treatments had a combination of inorganic and organic nutrients; and light treatments were obtained by covering the tanks with a single or double layer of screen. We measured esterase activity, oxidative stress (ROS), cell death, DNA damage, photosynthetic efficiency and 14 C assimilation as particulate or dissolved organic material (POC and DOC respectively). Conditions simulating future global change scenarios showed similar chlorophyllnormalized primary productivity as present conditions. The main effect driving phytoplankton physiology was the downregulation of the photosynthetic apparatus by elevated CO 2 , which decreased esterase activity, ROS, cell death and DNA damage. Nutrient concentration and light acted as additional modulators, upregulating or contributing to downregulation. The percentage of DO 14 C extracellular release (PER) was low (0 to 27%), significantly lower under ultraviolet radiation (UVR) than under photosynthetically active radiation (PAR), and acted mainly to reequilibrate the internal balance when cells grown under UVR were exposed to PAR. PER was almost 3 times lower under high CO 2 , confirming a higher resource use efficiency of phytoplankton under future CO 2 concentrations.
Antioxidant enzymes such as superoxide dismutase (SOD) play a key role in the removal of reactive oxygen species produced during visible and ultraviolet irradiance stress in microalgae and plants. However, little is known about the enzymatic antioxidative stress responses in ecologically important Antarctic marine microalgae. SOD in particular is difficult to analyze, possibly due to problems in obtaining sufficient quantities necessary for reliable and reproducible enzymatic assays. The aim of the present work was to create a sensitive, easy-to-use and reliable method for SOD determination in Antarctic microalgal material by comparing and optimizing existing protein extraction procedures and SOD assays in the marine Antarctic diatom Chaetoceros brevis. Optimization was achieved in cell disruption (sonication) and protein extraction procedures, extraction buffers, SOD assay methods (xanthine/xanthine oxidase and NBT/riboflavin photometric quantitative methods and native gel electrophoresis qualitative method) and the assay temperature. Protein extraction was optimal at low sonication amplitudes after a few pulses, irrespective of the type of buffer used. Extraction efficiency varied highly between the tested buffers; most protein was extracted in the presence of 1% of Triton X-100. SOD activity was best quantified using the NBT/riboflavin method in combination with a buffer containing potassium phosphate and Triton X-100. Moreover, the NBT/riboflavin method was demonstrated to be the most reliable and sensitive method at low temperatures (5 degrees C).
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