Virginia E. Villafañe

Photoacclimation of antarctic marine diatoms to solar ultraviolet radiation

Abstract The present study was carried out at Palmer Station (64.7 ° S, 64.1 ° W), Antarctica, during the austral spring-summer of the years 1993 and 1994. Two centric diatom species ( Thalassiosira sp. and Corethron criophilum Castracane) and two pennate species ( Pseudonitzschia sp. and Fragilariopsis cylindrus (Grunow) Krieger) were isolated from natural phytoplankton assemblages and exposed to solar radiation to study long term (more than 1 week) photoacclimation to ultraviolet radiation (UVR). At the beginning of the experiments, three of the cultures had relatively low concentrations of UV-absorbing compounds (i.e., mycosporine-like amino acids) and photosynthetic rates were significantly inhibited by UVR. At the end of the experiments (8–12 days), however, the two centric diatom species had high contents of mycosporine-like amino acids (MAAs) and did not show any significant differences in photosynthetic rates when exposed to either UVR + PAR or just to PAR. The synthesis of MAAs was slightly less when samples were exposed only to PAR than when exposed to UVR in addition to PAR. The rates of synthesis of MAAs, relative to phytoplankton carbon, for the two centric diatoms were 0.001 and 0.008 μg MAAs · (μg C) −1 · day −1 for shinorine and porphyra-334, respectively. The concentrations of MAAs in Pseudonitzschia sp., and Fragilariopsis cylindrus at the end of the experiments were much lower (less than one tenth) than that in the centric diatoms and the cultures were still inhibited by UVR. In the pennate diatoms MAAs increased in concentration as a response only to UVR and not to PAR. The loss rates of MAAs in Thalassiosira sp. after transferring the culture from high (1200 μE · m −2 · s −1 ) to low irradiance (250 μE · m −2 · s −1 ) were 0.0002 and 0.0023 μg MAAs · (μg C) −1 · day −1 for shinorine and porphyra-334, respectively. These results provide further evidence that MAA compounds are synthesized in response to high light conditions and that they do decrease the photoinhibitory effects of UVR.

Solar UV Radiation Drives CO2 Fixation in Marine Phytoplankton: A Double-Edged Sword

Photosynthesis by phytoplankton cells in aquatic environments contributes to more than 40% of the global primary production (Behrenfeld et al., 2006). Within the euphotic zone (down to 1% of surface photosynthetically active radiation [PAR]), cells are exposed not only to PAR (400-700 nm) but also to UV radiation (UVR; 280-400 nm) that can penetrate to considerable depths (Hargreaves, 2003). In contrast to PAR, which is energizing to photosynthesis, UVR is usually regarded as a stressor (Hader, 2003) and suggested to affect CO2-concentrating mechanisms in phytoplankton (Beardall et al., 2002). Solar UVR is known to reduce photosynthetic rates (Steemann Nielsen, 1964; Helbling et al., 2003), and damage cellular components such as D1 proteins (Sass et al., 1997) and DNA molecules (Buma et al., 2003). It can also decrease the growth (Villafane et al., 2003) and alter the rate of nutrient uptake (Fauchot et al., 2000) and the fatty acid composition (Goes et al., 1994) of phytoplankton. Recently, it has been found that natural levels of UVR can alter the morphology of the cyanobacterium Arthrospira (Spirulina) platensis (Wu et al., 2005b). On the other hand, positive effects of UVR, especially of UV- A (315-400 nm), have also been reported. UV- A enhances carbon fixation of phytoplankton under reduced (Nilawati et al., 1997; Barbieri et al., 2002) or fast-fluctuating (Helbling et al., 2003) solar irradiance and allows photorepair of UV- B-induced DNA damage (Buma et al., 2003). Furthermore, the presence of UV-A resulted in higher biomass production of A. platensis as compared to that under PAR alone (Wu et al., 2005a). Energy of UVR absorbed by the diatom Pseudo-nitzschia multiseries was found to cause fluorescence (Orellana et al., 2004). In addition, fluorescent pigments in corals and their algal symbiont are known to absorb UVR and play positive roles for the symbiotic photosynthesis and photoprotection (Schlichter et al., 1986; Salih et al., 2000). However, despite the positive effects that solar UVR may have on aquatic photosynthetic organisms, there is no direct evidence to what extent and howUVR per se is utilized by phytoplankton. In addition, estimations of aquatic biological production have been carried out in incubations considering only PAR (i. e. using UV-opaque vials made of glass or polycarbonate; Donk et al., 2001) without UVR being considered (Hein and Sand-Jensen, 1997; Schippers and Lurling, 2004). Here, we have found that UVR can act as an additional source of energy for photosynthesis in tropical marine phytoplankton, though it occasionally causes photoinhibition at high PAR levels. While UVR is usually thought of as damaging, our results indicate that UVR can enhance primary production of phytoplankton. Therefore, oceanic carbon fixation estimates may be underestimated by a large percentage if UVR is not taken into account.

Effects of solar UV radiation on morphology and photosynthesis of filamentous cyanobacterium Arthrospira platensis.

ABSTRACT To study the impact of solar UV radiation (UVR) (280 to 400 nm) on the filamentous cyanobacterium Arthrospira (Spirulina) platensis, we examined the morphological changes and photosynthetic performance using an indoor-grown strain (which had not been exposed to sunlight for decades) and an outdoor-grown strain (which had been grown under sunlight for decades) while they were cultured with three solar radiation treatments: PAB (photosynthetically active radiation [PAR] plus UVR; 280 to 700 nm), PA (PAR plus UV-A; 320 to 700 nm), and P (PAR only; 400 to 700 nm). Solar UVR broke the spiral filaments of A. platensis exposed to full solar radiation in short-term low-cell-density cultures. This breakage was observed after 2 h for the indoor strain but after 4 to 6 h for the outdoor strain. Filament breakage also occurred in the cultures exposed to PAR alone; however, the extent of breakage was less than that observed for filaments exposed to full solar radiation. The spiral filaments broke and compressed when high-cell-density cultures were exposed to full solar radiation during long-term experiments. When UV-B was screened off, the filaments initially broke, but they elongated and became loosely arranged later (i.e., there were fewer spirals per unit of filament length). When UVR was filtered out, the spiral structure hardly broke or became looser. Photosynthetic O2 evolution in the presence of UVR was significantly suppressed in the indoor strain compared to the outdoor strain. UVR-induced inhibition increased with exposure time, and it was significantly lower in the outdoor strain. The concentration of UV-absorbing compounds was low in both strains, and there was no significant change in the amount regardless of the radiation treatment, suggesting that these compounds were not effectively used as protection against solar UVR. Self-shading, on the other hand, produced by compression of the spirals over adaptive time scales, seems to play an important role in protecting this species against deleterious UVR. Our findings suggest that the increase in UV-B irradiance due to ozone depletion not only might affect photosynthesis but also might alter the morphological development of filamentous cyanobacteria during acclimation or over adaptive time scales.

Patterns of DNA damage and photoinhibition in temperate South-Atlantic picophytoplankton exposed to solar ultraviolet radiation

Natural marine phytoplankton assemblages from Bahía Bustamante (Chubut, Argentina, 45 degrees S, 66.5 degrees W), mainly consisting of cells in the picoplankton size range (0.2-2 microm), were exposed to various UVBR (280-315 nm) and UVAR (315-400 nm) regimes in order to follow wavelength-dependent patterns of cyclobutane pyrimidine dimer (CPD) induction and repair. Simultaneously, UVR induced photosynthetic inhibition was studied in radiocarbon incorporation experiments. Biological weighting functions (BWFs) for photoinhibition and for CPD induction, the latter measured in bare calf thymus DNA, differed in the UVAR region: carbon incorporation was reduced markedly due to UVAR, whereas no measurable UVAR effect was found on CPD formation. In contrast, BWFs for inhibition of photosynthesis and CPD accumulation were fairly similar in the UVBR region, especially above 300 nm. Incubation of phytoplankton under full solar radiation caused rapid CPD accumulation over the day, giving maximum damage levels exceeding 500 CPD MB(-1) at the end of the afternoon. A clear daily pattern of CPD accumulation was found, in keeping with the DNA effective dose measured by a DNA dosimeter. In contrast, UVBR induced photosynthetic inhibition was not dose related and remained nearly constant during the day. Screening of UVBR or UVR did not cause significant CPD removal, indicating that photoreactivation either by PAR or UVAR was of minor importance in these organisms. High CPD levels were found in situ early in the morning, which remained unaffected notwithstanding treatments favoring photorepair. These results imply that a proportion of cells had been killed by UVBR exposure prior to the treatments. Our data suggest that the limited potential for photoreactivation in picophytoplankton assemblages from the southern Atlantic Ocean causes high CPD accumulation as a result of UVBR exposure.

Impact of Solar Ultraviolet Radiation on Marine Phytoplankton of Patagonia, Argentina¶

Patagonia area is located in close proximity to the Antarctic ozone “hole” and thus receives enhanced ultraviolet B (UV‐B) radiation (280–315 nm) in addition to the normal levels of ultraviolet A (UV‐A; 315–400 nm) and photosynthetically available radiation (PAR; 400‐700 nm). In marine ecosystems of Patagonia, normal ultraviolet radiation (UVR) levels affect phytoplankton assemblages during the three phases of the annual succession: (1) prebloom season (late summer‐fall), (2) bloom season (winter‐early spring) and (3) postbloom season (late spring‐summer). Small‐size cells characterize the pre‐and postbloom communities, which have a relatively high photosynthetic inhibition because of high UVR levels during those seasons. During the bloom, characterized by micro‐plankton diatoms, photosynthetic inhibition is low because of the low UVR levels reaching the earths surface during winter; this community, however, is more sensitive to UV‐B when inhibition is normalized by irradiance (i.e. biological weighting functions). In situ studies have shown that UVR significantly affects not only photosynthesis but also the DNA molecule, but these negative effects are rapidly reduced in the water column because of the differential attenuation of solar radiation. UVR also affects photosynthesis versus irradiance (P vs E) parameters of some natural phytoplankton assemblages (i.e. during the pre‐ but not during the postbloom season). However, there is a significant temporal variability of P vs E parameters, which are influenced by the nutrient status of cells and taxonomic composition; taxonomic composition is in turn associated with the stratification conditions (e.g. wind speed and duration). In Patagonia, wind speed is one of the most important variables that conditions the development of the winter bloom by regulating the depth of the upper mixed layer (UML) and hence the mean irradiance received by cells. Studies on the interactive effects of UVR and mixing show that responses of phytoplankton vary according to the taxonomic composition and cell structure of assemblages; therefore cells use UVR if >90% of the euphotic zone is being mixed. In fact, cell size plays a very important role when estimating the impact of UVR on phytoplankton, with large cells being more sensitive when determining photosynthesis inhibition, whereas small cells are more sensitive to DNA damage. Finally, in long‐term experiments, it was determined that UVR can shape the diatom community structure in some assemblages of coastal waters, but it is virtually unknown how these changes affect the trophody‐namics of marine systems. Future studies should consider the combined effects of UVR on both phytoplankton and grazers to establish potential changes in biodiversity of the area.

Variability of UVR effects on photosynthesis of summer phytoplankton assemblages from a tropical coastal area of the South China Sea

From June to September 2005, we carried out experiments to determine the ultraviolet radiation (UVR) ‐induced photoinhibition of summer phytoplankton assemblages from a coastal site of the South China Sea. Variability in taxonomic composition was determined throughout the summer, with a peak chlorophyll a (chl a∼20 μg chl a L−1) dominated by the diatom Skeletonema costatum that was detected early in the study period; the rest of the time samples were characterized by monads and flagellates, with low chl a values (1–5 chl a μg L−1). Surface water samples were placed in quartz tubes, inoculated with radiocarbon and exposed to solar radiation for 2–3 h to determine photosynthetic rates under three quality radiation treatments (i.e. PAB, 280–700 nm; PA, 320–700 nm and P, 400–700 nm) using different filters and under seven levels of ambient irradiance using neutral density screens (PvsE curves). UVR inhibition of samples exposed to maximum irradiance (i.e. at the surface) varied from −12.2% to 50%, while the daytime‐integrated UVR‐related photoinhibition in surface seawater varied from −62% to 7%. The effects of UVR on the photosynthetic parameters PBmax and Ek were also variable, but UV‐B accounted for most of the observed variability. During sunny days, photosynthesis of microplankton (>20 μm) and piconanoplankton (<20 μm) were significantly inhibited by UVR (mostly by UV‐B). However, during cloudy days, while piconanoplankton cells were still inhibited by UVR, microplankton cells used UVR (mostly UV‐A) as the source of energy for photosynthesis, resulting in higher carbon fixation in samples exposed to UVR than the ones exposed only to photosynthetically active radiation (PAR). Our results indicate that size structure and cloudiness clearly condition the overall impact of UVR on phytoplankton photosynthesis in this tropical site of South China. In addition, model predictions for this area considering only PAR for primary production might have underestimated carbon fixation due to UVR contribution.

DNA damage and photosynthetic inhibition induced by solar ultraviolet radiation in tropical phytoplankton (Lake Titicaca, Bolivia)

Experiments were conducted during October 1998 in Lake Titicaca, Bolivia (16° S, 68° W, 3810 m a.s.l), to determine the effects of solar ultraviolet radiation (UVR) on phytoplankton photosynthetic rates and DNA damage. Water samples were taken daily and incubated in situ or in simulated in situ conditions using sharp cut-off filters to eliminate various portions of the UVR spectrum. The total inhibition of photosynthesis due to UVR in surface waters was 85%; the greatest part of this inhibition (65%) was due to UVAR (315–400 nm), the rest (20%) being due to UVBR (280–315 nm). The inhibition of photosynthesis decreased with depth so that there were no significant differences among treatments at 1.3 optical depths (K PAR). The loss of carbon assimilation in the integrated production over the euphotic zone (4.6 optical depths) was 17.4%, with 14% due to UVAR and an additional 3.4% due to UVBR. Lake Titicaca phytoplankton had a reshold for inhibition of photosynthesis at about 0.3 W m−2 for UVBR and 5 W m−2 for UVAR, below which no inhibition was detected. Above this threshold, photosynthetic inhibition increased steadily, with UVAR having the greatest effect. Analysis of biological weighting functions (BWFs) indicated that phytoplankton from Lake Titicaca was less sensitive to UVR than phytoplankton from other regions. DNA damage (evaluated through the formation of cyclobutane pyrimidine dimers, CPDs) was observed for a simulated worst-case situation (i.e. samples incubated in full sunlight) and significant CPD accumulation was found as a result of UVBR exposure, but not with UVAR. However, absolute levels of damage were relatively low when compared with results obtained at other locations, also suggesting the low sensitivity of Lake Titicaca phytoplankton to UVR. It seems that UVBR stress in these organisms acts via at least two cellular targets: the photosynthetic apparatus and nuclear DNA. Our results suggest that an eventual enhancement of UVBR, due to ozone depletion, would have little impact on the phytoplankton of Lake Titicaca.

Combined effects of solar ultraviolet radiation and nutrients addition on growth, biomass and taxonomic composition of coastal marine phytoplankton communities of Patagonia

Experiments (6-8 days) were conducted during late summer, late fall and late winter, 2003 with waters collected off Bahía Nueva, Chubut, Argentina (42.7 degrees S, 65 degrees W) to determine the combined effects of solar ultraviolet radiation (UVR, 280-400 nm) and nutrient addition on phytoplankton communities. Samples were put in UVR-transparent containers and incubated under two radiation treatments: (a) Samples exposed to full solar radiation (PAB treatment, 280-400 nm) and (b) Samples exposed only to PAR (PAR treatment, 400-700 nm). At the beginning of the experiments, nutrients (i.e., NaPO(4)H(2) and NaNO(3)) were added to one set of samples from each radiation treatment (N cultures) whereas in the other set, nutrients remained at the concentration of the seawater. Chlorophyll a, biomass, UV-absorbing compounds and taxonomic composition were recorded throughout the experiments. N cultures always had significantly higher growth rates (P<0.05) than that in non-enriched cultures. At the beginning of experiments, phytoplankton communities were generally dominated by monads and flagellates but by the end, diatoms comprised the bulk of biomass, with only one to four taxa dominating, suggesting a selection towards more tolerant/less sensitive species. Over the experimental time frame, the observed taxonomic changes were mostly due to nutrient availability, and to a lesser extent to solar UVR exposure. Overall, the results indicate that environmental conditions (i.e., light history, nutrient concentration) together with the physiological status of the cells play a very important role at the time to assess the combined effect of nutrient addition and solar radiation on coastal phytoplankton assemblages from Patagonia.

Bioaccumulation and role of UV-absorbing compounds in two marine crustacean species from Patagonia, Argentina

Experiments were conducted during summer and winter, 2000, and summer 2001 to determine the bioaccumulation and role of UV-absorbing compounds in two crustacean species--the amphipod Amphitoe valida and the isopod Idothea haltica--from the mid-littoral of the Patagonia coast (Argentina). Macroalgae constituting the diet for these species differed in the concentration of UV-absorbing compounds, from high amounts in the rhodophyte Polysiphonia sp. to almost null in chlorophyte species (i.e., Enteromorpha sp. and Codium sp.). Consequently, transferring and bioaccumulation of these compounds, identified as the mycosporine-like amino acids (MAAs) Porphyra-334 and Shinorine, varied in the crustaceans according to their algal diet, being high when feeding on Polysiphonia sp. Survival experiments carried out with crustaceans feeding on poor and rich-MAA diets demonstrated that the role of these compounds in A. valida and I. baltica was different. In A. valida, and based on a significantly higher survival in those individuals feeding on the rhodophyte, MAAs seem to provide an effective protection against UV-B radiation (280-320 nm). In I. baltica, mortality was not significantly different in individuals feeding on rich and poor MAA diets. However, judging from the comparatively high amounts of MAAs in eggs/embryos, these compounds might provide protection to the progeny rather than to adult organisms.

Temperature benefits the photosynthetic performance of the diatoms Chaetoceros gracilis and Thalassiosira weissflogii when exposed to UVR.

The aim of this study was to assess the combined effects of temperature and UVR on the photosynthesis performance of two diatoms -Chaetoceros gracilis and Thalassiosira weissflogii. In particular, we evaluated the role of UVR in inducing photoinhibition and the potential mitigation of this negative effect by an increase in temperature. Cultures were pre-acclimated at two temperatures - 18°C and 23°C - and exposed to different radiation treatments - UVR+PAR (280-700nm); UV-A+PAR (315-700nm) and PAR only (400-700nm) under two temperatures: 18°C (local surface summer water temperature) and 23°C (simulating a potential increase estimated by the year 2100). Exposure to natural solar radiation resulted in UVR-induced photoinhibition that was significantly higher in T. weissflogii than in C. gracilis. Both species benefited from the higher temperature (23°C) resulting in a lower photoinhibition as compared to samples exposed at 18°C. Inter-specific differences were determined in regard to the heat dissipation processes (NPQ) which were higher at high temperatures, and much more evident in C. gracilis than in T. weissflogii. The analyses of inhibition and recovery rates under different irradiances indicate that the balance between negative (inhibition) and positive (repair-dissipation) effects shifted towards a more positive balance with increasing temperature. Our results highlight for a beneficial effect of temperature on photosynthesis performance during exposure to UVR, although important inter-specific differences are found, probably due to differences in cell size as well as in their distribution within the oceanic realm (i.e., coastal versus oceanic species).