Diurnal measurements of F v/F m used to improve productivity estimates in macroalgae

The ratio of variable fluorescence to maximal fluorescence (F _v/F _m) was measured during a night and day cycle in five different macroalgae growing in the littoral zone at the Swedish west coast; the green algae Ulva lactuca, Cladophora sp. and Enteromorpha flexousa, the red alga Ceramium nodulosum and the brown alga Fucus vesiculosus. All the green algae and C. nodulosum showed both diurnal fluctuations in F _v/F _m during days of high light intensities and a significant negative correlation between F _v/F _m and photon irradiance (PI). An attempt has been made to improve calculations of macroalgal net growth based on carbon fixation (α_net) considering this diurnal change in F _v/F _m. By assuming that the negative linear correlation between F _v/F _m and the maximum quantum yield for photosynthesis (Φ) is proportional to that between F _v/F _m and α_net, it was possible to include the daily variation of α_net due to photoinhibition. To compensate for the variation of F _v/F _m, a representative value for each day was obtained by weighting all values of F _v/F _m over the diurnal period in relation to total PI. For all algal species there was a fairly good agreement between this representative value and the F _v/F _m value measured around noon. As the daily representative F _v/F _m value showed a negative linear correlation with the daily mean PI, it was possible to correct α_net for differences of daily mean PI. Received: 10 March 1997 / Accepted: 25 August 1997     

Evaluation of chlorophyll fluorescence, in vivo spectrophotometric pigment absorption and ion leakage as biomarkers of UV-B exposure in marine macroalgae

The photosynthetic fluorescence ratio F_v:F_m, in vivo absorption spectra and ion leakage were evaluated as biomarkers of ambient and elevated UV-B (280 to 320 nm) exposure of the intertidal alga Enteromorpha intestinalis (Chlorophyta) and the sublittoral alga Palmaria palmata (Rhodophyta). Measurements of thallus growth were also used to assess adverse biological effects. Ambient and elevated UV-B significantly inhibited photosynthesis in both species. It was shown that the F_v:F_m ratio is a sensitive, non-specific general biomarker of UV-B exposure in both species. Moreover, the in vivo absorption of what was tentatively identified as chlorophylls a and b as well as phycoerythrin and/or carotenoids, phycoerythrobilin and phycocyanin decreased in a dose-response dependent manner and was associated with a decrease in growth rate in P. palmata. The intertidal alga E. intestinalis showed a greater degree of tolerance to UV-B exposure. These results indicate that changes in the F_v:F_m ratio together with reductions in in vivo pigment absorption could provide an early quantitative warning of the detrimental effects of UV-B in marine macroalgae. Received: 16 May 1997 / Accepted: 16 July 1997     

The role of the amphipod <Emphasis Type="Italic">Gammarus locusta</Emphasis> as a grazer on macroalgae in Swedish seagrass meadows

Mesograzers are thought to play a critical role in seagrass beds by preventing overgrowth of ephemeral algae. On the Swedish west coast, eelgrass Zostera marina has decreased in recent decades as a result of eutrophication and increased growth of macroalgal mats (mainly filamentous Ulva spp. and Ectocarpales), with no indication of grazer control of the algae. The aim of this study was to investigate the ability of the amphipod Gammarus locusta to control algal blooms during nutrient-enriched and ambient conditions, using a combination of laboratory, field and model studies. Laboratory experiments demonstrated that juvenile and adult G. locusta could consume both Ulva spp. and Ectocarpales, but that consumption of Ulva spp. was significantly higher. Cannibalism was common in individual treatments involving multiple size-classes of G. locusta, but only large, male gammarids consumed smaller juveniles in the presence of Ulva spp. as an alternative food source. However, no negative effects of cannibalism were found on total grazing impact. A model using size-specific grazing rates and growth rates of G. locusta and of Ulva spp. suggests that approximately 62 young juvenile, or 27 adult G. locusta are needed per gram DW of Ulva spp. to control the algal growth during ambient nutrient conditions, and approximately 2.6 times as many gammarids during enhanced nutrient conditions. On the Swedish west coast, densities and mean sizes of G. locusta in eelgrass beds are below these critical values, suggesting that the gammarids will not be able to control the growth of the filamentous macroalgae. However, in the field cage experiment, immigration of juveniles and reproduction of encaged adult G. locusta resulted in unexpectedly high densities of G. locusta (>4,000 individual m⁻²), and very low biomass of Ulva spp. in both ambient and nutrient-enriched treatments. Although the high numbers of juveniles in all cages precluded any significant treatment effects, this suggests that in the absent of predators, the population of G. locusta can grow significantly and control the biomass of Ulva spp. Furthermore, low grazing of Ectocarpales in the laboratory and high biomass of these filamentous brown algae in the field indicate a preference for the more palatable green algae Ulva spp. This study indicates that the high grazing capacity of G. locusta, in combination with high reproduction and growth rates, would allow the amphipod to play a key role in Z. marina ecosystems by controlling destructive blooms of filamentous green algae. However, high predation pressure appears to prevent large populations of G. locusta in eelgrass beds on the Swedish west coast today.     

Inhibition of larval settlement to a soft bottom benthic community by drifting algal mats: An experimental test

The hypothesis that drifting red algal mats inhibit settlement of planktonic larvae was tested in a field experiment in 1986. Substratum free of algae (caged boxes) was compared with substratum covered with algae (natural substratum and open boxes). Whereas settling densities of 1500 to 5500 ind. m^-2 of the bivalves Macoma balthica, Cardium glaucum and Mya arenaria were observed in sediment without algal cover, no recruits of the same species were found beneath the algae during the period of peak settlement (June to July). The same difference was observed for the polychaete Nereis diversicolor, although in this case occasional individuals were found beneath the algae. The results demonstrate that algae mats may be efficient inhibitors of larval settlement to benthic soft-substratum communities. It is concluded that algae act as a larval filter.     

Size-dependent photosynthetic performance in the giant clam <Emphasis Type="Italic">Tridacna maxima</Emphasis>, a mixotrophic marine bivalve

Giant clams form a symbiosis with photosynthetic algae of the genus Symbiodinium that reside in clam mantle tissue. The allometry of symbiont photosynthetic performance was investigated as a mechanism for the increasing percentage of giant clam carbon respiratory requirements provided by symbionts as clam size increases. Chlorophyll fluorescence measurements of symbionts of the giant clam Tridacna maxima were measured during experiments conducted in September of 2009 using specimens 0.5–200 g tissue wet weight (3–25 cm long), collected from waters around southern Taiwan (N 21°36′, E 120°47′) from July to August of 2009. Light-dependent decreases in effective quantum yield (∆F/F _m′) calculated as the noontime maximum excitation pressure over PSII (Q _m), relative electron transport rates (rETR), and dark-adapted maximum quantum yield (F _v/F _m) all varied as a quadratic function of clam size. Both Q _m and rETR increased as clam size increased up to ~10–50 g then decreased as clam size increased. F _v/F _m decreased as clam size increased up to ~5–50 g then increased as clam size increased. Chlorophyll fluorescence measurements of rETR were positively correlated with gross primary production measured during chamber incubations. Overall, symbionts of mid-sized clams ~5–50 g exhibited the highest light-dependent decreases in effective photosynthetic efficiencies, the highest relative electron transport rates, and the lowest maximum photosynthetic efficiencies, and symbiont photosynthetic performance is allometric with respect to host clam size.     

Interactive effects of UV radiation and enhanced temperature on photosynthesis, phlorotannin induction and antioxidant activities of two sub-Antarctic brown algae

Lessonia nigrescens and Durvillaea antarctica, two large sub-Antarctic brown algae from the southern Chilean coast, were exposed to solar UV radiation in an outdoor system during a summer day (for 11 h) as well as to artificial UV radiation under controlled laboratory conditions at two temperatures (15 and 20 °C) for 72 h. Chlorophyll a fluorescence–based photoinhibition of photosynthesis was measured during the outdoor exposure, while electron transport rates, lipid peroxidation, antioxidant activity and content of phlorotannins were determined at different time intervals during the laboratory exposure. Under natural solar irradiances in summer, both species displayed well-developed dynamic photoinhibition: F _v/F _m values decreased by 70 % at noon coinciding with the levels of PAR >1,500 μmol m^?2 s^?1 and UV-B radiation >1 W m^?2 and recovered substantially in the afternoon. In treatments including UV radiation, recovery in D. antarctica started already during the highest irradiances at noon. The results from laboratory exposures revealed that (a) elevated temperature of 20 °C exacerbated the detrimental effects of UV radiation on photochemical parameters (F _v/F _m and ETR); (b) peroxidative damage measured as MDA formation occurred rapidly and was strongly correlated with the decrease in F _v/F _m, especially at elevated temperature of 20 °C; (c) the antioxidant activity and increases in soluble phlorotannins were positively correlated mainly in response to UV radiation; (d) phlorotannins were rapidly induced but strongly impaired at 20 °C. In general, short-term (2–6 h) exposures to enhanced UV radiation and temperature were effective to activate the photochemical and biochemical defenses against oxidative stress, and they continued operative during 72 h, a time span clearly exceeding the tidal or diurnal period. Furthermore, when algae were exposed to dim light and control temperature of 15 °C for 6 h, F _v/F _m increased and lipid peroxidation decreased, indicating consistently that algae retained their ability for recovery. D. antarctica was the most sensitive species to elevated temperature for prolonged periods in the laboratory. Although no conclusive evidence for the effect of the buoyancy of fronds was found, the interspecific discrepancies in thermo-sensitivity in the UV responses found in this study are consistent with various ecological and biogeographical differences described for these species.     

Effect of hyposalinity on the photophysiology of <Emphasis Type="Italic">Siderastrea radians</Emphasis>

Tolerance to hyposalinity of the scleractinian coral S. radians was examined in a mesocosm study. Colonies of S. radians were collected from five basins in Florida Bay, USA, which occur along a northeast-to-southwest salinity gradient. Salinity treatments were based on historical salinity records for these basins. Photophysiology of the endosymbiont Symbiodinium spp. (maximum quantum yield; F _v/F _m) was measured as an indicator of holobiont stress to hyposalinity. Colonies from each basin were assigned four salinity treatments [The Practical Salinity Scale (PSS) was used to determine salinity. Units are not assigned to salinity values because it is a ratio and has no unit as defined by UNESCO (UNESCO Technical papers no. 45, IAPSO Pub. Sci. No. 32, Paris, France, 1985)] (30, 20, 15, and 10) and salinities were reduced 2 per day from ambient (30) to simulate a natural salinity decrease. Colonies treated with salinities of 20 and 15 showed no decrease in F _v/F _m versus controls (i.e. 30), up to 5 days after reaching their target salinity. This indicates a greater ability to withstand reduced salinity for relatively extended periods of time in S. radians compared to other reef species. Within 1 day after salinity of 10 was reached, there was a significant reduction in F _v/F _m, indicating a critical threshold for hyposaline tolerance. At the lowest treatment salinity (10), F _v/F _m for the more estuarine, northeast-basin colonies were significantly higher than the most marine southwest-basin colonies (Twin Key Basin). Our results suggest that historical salinity ranges within basins determine coral population salinity tolerances.     

Food preferences and related behavior of the browsing sea urchin <Emphasis Type="Italic">Tripneustes gratilla</Emphasis> (Linnaeus) and its potential for use as a biological control agent

The short-spined toxopneustid sea urchin Tripneustes gratilla feeds on a wide variety of algal species and on sea grasses. However, the urchin does show preferences when offered a selection of macroalgal species, which it encounters in nature. Preferences among macroalgae were evident in field-collected urchins exposed to pair-wise tests where the variable was either the consumption rate of the algae or observation of which algal species the urchins chose to touch with their lantern teeth. Exposure of lab-housed urchins to one of five species of macroalgae for 5 months did not seem to alter preferences of urchins in three of the exposure groups, but those exposed to Padina sanctae-crucis seemed to show an enhanced preference for this species when offered a choice of the five species of macroalgae at the end of the exposure period, and those exposed to Gracilaria salicornia seemed to avoid the species when offered the choice of the five species. Perhaps more ecologically important than their preferences were two other observations on these urchins: first, when offered only a single species of algae, the urchins on four of five diets ate the same quantity per day. Second, when simultaneously offered the choice among the five macroalgal species, the urchins consumed more macroalgae per day than when offered only one species. These urchins move about a meter a day. They probably encounter food resources in a relatively coarse-grained fashion and have evolved to eat what is available. Because of their limited movements, their habitat overlap with grazing fishes, their acceptance of a wide variety of macroalgae and their preference for macroalgae, these native urchins are thought to have the potential to serve as biological control agents of alien and invasive macroalgae, which have come to dominate some reef zones normally occupied by corals in Hawaii.     

Response of zooxanthellae in symbiosis with the Mediterranean corals Cladocora caespitosa and Oculina patagonica to elevated temperatures

Scleractinian symbiotic corals living in the Ligurian Sea (NW Mediterranean Sea) have experienced warm summers during the last decade, with temperatures rapidly increasing, within a few days, to 3–4°C above the mean value of 24°C. The effect of elevated temperatures on the photosynthetic efficiency of zooxanthellae in symbiosis with temperate corals has not been well investigated. In this study, the corals, Cladocora caespitosa and Oculina patagonica were collected in the Ligurian Sea (44°N, 9°E), maintained during 2 weeks at the mean summer temperature of 24°C and then exposed during 48 h to temperatures of 24 (control), 27, 29 and 32°C. Chlorophyll (chl) fluorescence parameters [F _v/F _m, electron transport rate (ETR), non-photochemical quenching (NPQ)] were measured using pulse amplitude modulated (PAM) fluorimetry before, during the thermal increase, and after 1 and 7 days of recovery (corals maintained at 24°C). Zooxanthellae showed a broad tolerance to temperature increase, since their density remained unchanged and there was no significant reduction in their maximum quantum yield (F _v/F _m) or ETR up to 29°C. This temperature corresponded to a 5°C increase compared to the mean summer temperature (24°C) in the Ligurian Sea. At 32°C, there was a significant decrease in chl contents for both corals. This decrease was due to a reduction in the chl/zooxanthellae content. For C. caespitosa, there was also a decrease in ETR_max, not associated with a change in F _v/F _m or in the non-photochemical quenching (NPQ); for O. patagonica, both ETR_max and F _v/F _m significantly decreased, and NPQ_max showed a significant increase. Damages to the photosystem II appeared to be reversible in both corals, since F _v/F _m values returned to normal after 1 day at 24°C. Zooxanthellae in symbiosis with the Mediterranean corals investigated can therefore be considered as resistant to short-term increases in temperature, even well above the maximum temperatures experienced by these corals in summer.     

Selective consumption and facilitation by mesograzers in adult and colonizing macroalgal assemblages

Grazing pressure on macroalgae in littoral communities may vary with algal species, the age of an algal individual and grazer identity. Previous studies on alga–grazer interactions have shown that grazer preference for an algal species may release another one from interspecific competition. We measured the impacts of four common grazer taxa and the natural grazer guild on macroalgal communities at both their colonization and adult stages, and compared the impacts to grazer exclosures. The grazer effects were stronger on colonizing than on adult macroalgae; grazers did not reduce the total density of adult algae. Grazers both feed on propagules and indirectly facilitate other algae, depending on the grazer or algal species. Hydrobia species increased the settlement of spores of the red alga Ceramium tenuicorne. Similarly, the gastropod Theodoxus fluviatilis tended to facilitate one crustose algal species, but decreased the propagule density of annual filamentous algae, suggesting a preference for one species to the advantage of another. Effects of crustacean mesograzers on the studied macroalgae were weak. These results indicate that northern Baltic macroalgae are limited to grazing mainly during their colonization stage.     

Effects of light exposure on the retention of kleptoplastic photosynthetic activity in the sacoglossan mollusc <Emphasis Type="Italic">Elysia viridis</Emphasis>

The effects of light exposure on the photosynthetic activity of kleptoplasts were studied in the sacoglossan mollusc Elysia viridis. The photosynthetic activity of ingested chloroplasts was assessed in vivo by non-destructively measuring photophysiological parameters using pulse amplitude modulation (PAM) fluorometry. Animals kept under starvation were exposed to two contrasting light conditions, 30 μmol photons m⁻² s⁻¹ (low light, LL), and 140 μmol photons m⁻² s⁻¹ (high light, HL), and changes in photosynthetic activity were monitored by measuring the maximum quantum yield of photosystem II (PSII), F _v/F _m, the minimum fluorescence, F _o, related to chlorophyll a content, and by measuring rapid light-response curves (RLC) of relative electron transport rate (rETR). RLCs were characterised by the initial slope of the curve, α_RLC, related to efficiency of light capture, and the maximum rETR level, rETR_m,RLC, determined by the carbon-fixation metabolism. Starvation induced the decrease of all photophysiological parameters. However, the retention of photosynthetic activity (number of days for F _v/F _m > 0), as well as the rate and the patterns of its decrease over time, varied markedly with light exposure. Under HL conditions, a rapid, exponential decrease was observed for F _v/F _m, α_RLC and rETR_m,RLC, F _o not showing any consistent trend of variation, and retention times ranged between 6 and 15 days. These results suggested that the retention of chloroplast functionality is limited by photoinactivation of PSII reaction center protein D1. In contrast, under LL conditions, a slower decrease in all parameters was found, with retention times varying from 15 to 57 days. F _v/F _m, α_RLC and rETR_m,RLC exhibited a bi-phasic pattern composed by a long phase of slow decrease in values followed by a rapid decline, whilst F _o decayed exponentially. These results were interpreted as resulting from lower rates of D1 photoinactivation under low light and from the gradual decrease in carbon provided by photosynthesis due to reduction of functional photosynthetic units.     

Capability of dynamic photoinhibition in Arctic macroalgae is related to their depth distribution

The capability of several macroalgal species to protect photosynthesis from excessive irradiance by dynamic photoinhibition was investigated relative to their depth distribution in summer 1995 in the Kongsfjord (79°N; 12°E, Ny Ålesund, Spitsbergen, Norway). Photoinhibition of photosynthesis was induced by exposure of algae from different water depths to a high photon fluence rate of 500?μmol?m^?2?s^?1 for 2?h. Changes in optimal quantum yield (F _v/F _m) were measured during the inhibition phase. Recovery of photosynthesis was subsequently induced by dim white light (10?μmol?m^?2?s^?1) and observed as changes in the variable fluorescence. With a newly developed mathematical model different parameters of the response kinetics of inhibition and recovery were calculated and related to the depth distribution of each algal species. It is shown that two components with slow and fast reaction kinetics, respectively, are involved in photoinhibition and recovery of photosynthesis. Their possible molecular bases are discussed. The half-life time (τ) of the inhibition and recovery phases, i.e. the time necessary to reach half maximal response, is clearly related to the depth distribution of the investigated species. Algae collected close to the water surface show a fast reaction of both photoinhibition and recovery and, hence, have a low τ. With increasing depth the reactions become slower and τ increases. τ was highest in deep water algae. Further analysis of the reaction kinetics in Laminaria saccharina shows that the relative proportion of the two kinetics involved change with the collection depth. In contrast, a significant difference in the reaction rates of both kinetics was not observed.     

Influence of ultraviolet-radiation on chlorophyll fluorescence and growth in different life-history stages of three species of Laminaria (phaeophyta)

Zoospores, gametophytes, young sporophytes and discs cut from mature sporophytes of Laminaria digitata, L. hyperborea and L. saccharina were exposed in the laboratory to UV-radiation, with a spectral composition and irradiance similar to natural sunlight, for periods ranging from 15 min to 8 d, and were then returned to white light. Germination of zoospores and the growth of gametophytes were reduced after exposures to UV longer than 1 h, whereas UV had little effect on the growth of young or mature sporophytes unless exposure continued for more than 48 h. The variable fluorescence (F _v:F_m) of all stages was strongly reduced immediately after short exposures to UV, but recovered almost completely within 24 h. However, exposure of gametophytes to UV for >4 h resulted in little or no recovery of F _v:F_m, whereas >16 h of UV were required to produce this result in young sporophytes, and >48 h in mature sporophytes. Thus, sensitivity to UV-radiation decreased from gametophytes to sporophytes, and with increasing age of sporophytes, but, in gametophytes, growth appeared to be a more sensitive indicator of UV-damage than F _v:F_m after 24 h recovery. The responses to UV of the zoospores and gametophytes of all three species were similar, but both growth and fluorescence measurements suggested that the sporophytes of L. saccharina were more sensitive to UV than those of the other two species.     

Salinity stress and hydrogen peroxide regulation of antioxidant defense system in Ulva fasciata

The regulation of antioxidant defense system in macroalgae exposed to salinity stress was examined in Ulva fasciata Delile. As compared to the 30‰ control, a long-term (4 days) exposure to hyposaline (5, 15‰) and hypersaline (60, 90, 120, 150‰) conditions inhibited growth rate and TTC reduction ability. A decrease in maximum quantum efficiency (F _v/F _m ratio) and the maintenance of superoxide dismutase activity under salinity stress indicate the potential generation of reactive oxygen species in chloroplasts. An exposure to 15, 60, and 90‰ decreased seawater H₂O₂ contents but increased thallus H₂O₂ contents that are positively correlated with TBARS and peroxide contents. Alleviation of oxidative damage and H₂O₂ accumulation at 15 and 90‰ by a H₂O₂ scavenger, dimethylthiourea, suggests that oxidative damage occurring under moderate hyposaline and hypersaline conditions is ascribed to accumulated H₂O₂. Increased glutathione reductase activity and glutathione content and decreased ascorbate content are responsible for accumulated H₂O₂ at 15, 60, and 90‰, while ascorbate peroxidase activity increased only at salinity ≥ 90‰. Catalase and peroxidase activities also increased at 60 and 90‰ for H₂O₂ removal, but only catalase showed activity increase at 15‰. For the regeneration of ascorbate, the activities of both dehydroascorbate reductase and monodehydroascorbate reductase were increased at 5 and 15‰ while only monodehydroascorbate reductase activity increased at 60 and 90‰. It is hypothesized that the availability of antioxidants and the activities of antioxidant enzymes are increased in U. fasciata to cope with the oxidative stress occurring in hyposaline and hypersaline conditions.     

Photoinhibition and recovery of the kelp Laminaria saccharina at optimal and superoptimal temperatures

Effects of high irradiance on photosynthetic characteristics were examined in sporophytes of the kelp Laminaria saccharina Lamour. from 1992 to 1994. Exposure to high irradiance (700 mol photons m^-2s^-1) for 1 h at optimal temperature (12°C) caused a 40 to 60% decline in photosynthetic efficiency (alpha), quantum yield, and the ratio of variable to maximum chlorophyll fluorescence (F_v/F_m), an indicator of Photosystem II efficiency. Although the photoinhibition effects were partly attributable to protective mechanisms, a concurrent increase in minimal fluorescence (F_o) indicated damage to Photosystem II reaction centers. The magnitude of photoinhibition was proportional to irradiance and duration; however, F_v/F_m was significantly reduced after exposure to irradiances as low as 40 to 50 mol photons m^-2s^-1 for 1 h, or to 700 mol photons m^-2s^-1 for only 5 min. In contrast, photosynthetic capacity (P_max) was affected only at much higher irradiance. Superoptimal temperatures up to 24°C did not exacerbate high-light effects. At 25°C, however, alpha and P_max were more susceptible to photoinhibition than at lower temperatures. Recovery from photoinhibition was examined by following F_v/F_m and F_o for 24 h after exposure to high light. Recovery of F_v/F_m was fastest during the first 1 to 3 h, and slowed or ceased after 6 to 12 h, while recovery of F_o was relatively constant over 12 h. Dithiothreitol, which blocks formation of energy-dissipating xanthophylls, reduced both the initial rate and extent of recovery. Chloramphenicol, which blocks chloroplast-encoded protein synthesis, had little effect on initial rates of recovery, but stopped recovery after 3 h. Thus, L. saccharina appears to rely on the xanthophyll cycle to protect the photosynthetic apparatus, and reversal of this protective mechanism causes the rapid initial recovery in F_v/F_m. Longterm recovery depends on repair of damaged reaction centers. Both the rate and extent of recovery were temperature-dependent. The initial rate was higher at 18 to 22°C than at 12°C, but the extent of recovery over 24 h declined with increasing temperature. High temperatures, therefore, appear to enhance protective mechanisms, but disrupt repair processes. L. saccharina from Long Island Sound, an ecotype adapted to low light and high temperature, showed slightly but consistently greater effects of photoinhibition than plants from the Atlantic coast of Maine, but exhibited faster recovery at superoptimal temperatures.     

Characterization of in vivo absorption features of chlorophyte,phaeophyte and rhodophyte algal species

Despite the plentiful diversity of macroalgae in coastal environments, few studies have examined the in vivo absorption features of common marine macrophytes. Here we report on results of a survey of 12 central California common intertidal and subtidal taxa, representing Chlorophyta, Phaeophyta and Rhodophyta carried out in the summer of 1988. Computer-assisted analyses were used to obtain fourth-derivative spectra from in vivo absorption spectra determined at room temperature to obtain spectral diagnostics for the different algal divisions and to provide a means to determine whether spectral features could be used to identify stress responses among these plants. Among the Chlorophyta, characteristic maxima for chlorophylls a and b were resolved in all species examined, and a spectral component attributable to siphonaxanthin-like carotenoid(s) was observed in two Ulva species and the coenocytic alga Derbesia marina. Representatives of the Phaeophyta were characterized by similar maxima for chlorophyll a and c, and for fucoxanthin. Among the Rhodophyta, maxima for chlorophyll a, as well as B- and R-type phycoerythrin were resolved. Differences in in vivo absorption features were detected for two tidal populations of Porphyra perforata and Mastocarpus papillatus. High-tidal thalli absorb less green light in regions characterized by phycoerythrin, but have enhanced carotenoid absorption compared with lowtidal thalli. Resolution of spectra by fourth-derivative analysis revealed significant differences in phycoerythrin and carotenoid contents. The spectral changes observed appear to reflect environmental and possibly populational characteristics of these algae. The spectral analyses described here provide robust, non-invasive means to characterize subtle responses of macroalgae to environment in ways not possible previously. Additional merits of these fourth-derivative analyses for use in environmental studies are discussed.This contribution is dedicated to the memory of Professor L. R. Blinks, a pioneer in photosynthesis research and algal physiology     

Availability of ammonium influences photosynthesis and the accumulation of mycosporine-like amino acids in two<Emphasis Type="Italic"> Porphyra</Emphasis> species (Bangiales,Rhodophyta)

The effect of ammonium concentration on photosynthetic activity estimated as in vivo chlorophyll fluorescence, i.e. maximal quantum yield (F_v/F_m) and electron transport rate (ETR) and on the accumulation of mycosporine-like amino acids (MAAs), chlorophyll a (chl a), biliproteins (BP) and soluble proteins (SP) in the red algae Porphyra leucosticta Thuret in Le Jolis collected from Lagos (Málaga, Spain) and Porphyra umbilicalis (Linnaeus) J. Agardh from Helgoland (Germany) was evaluated. Discs of both species were incubated with three ammonium concentrations (0, 100 and 300 µM) under artificial PAR and UV radiation for 7 days. Photosynthetic activity decreased under the culture conditions due to UV radiation and ammonium availability. The decrease of both F_v/F_m and maximal ETR was related to ammonium supply, i.e. the lowest decrease occurred in algae growing with the highest concentration of ammonium. In both species, after 7 days of culture, the content of chl a, BP and SP was higher under 300 µM than that under 0 and 100 µM ammonium. In both species, the content of MAAs was increased under 300 µM ammonium compared to the initial value, whereas a decrease under 0 and 100 µM ammonium was observed only in P. leucosticta. The content of MAAs in P. umbilicalis did not present significant differences compared to the initial value, probably because of the high initial content of MAAs. In both Porphyra species, four MAAs were identified: shinorine, porphyra-334, palythine and asterina-330. However, P. leucosticta modified its MAA pattern during the incubation time, reaching the same percentages found for P. umbilicalis, which did not show any change during the experimental period. P. leucosticta exhibited a decrease in BP/SP and BP/chl a ratios through the incubation time and an increase in MAAs/BP. The ratio MAAs/chl a did not show any variation with time or treatment, as was also true for all ratios in P. umbilicalis. In summary, ammonium supply diminished the decrease of F_v/F_m, increased the content of photosynthetic pigments (chlorophyll and biliprotein) and soluble protein, and stimulated of the accumulation of MAAs in the red algae P. leucosticta and P. umbilicalis.Communicated by O. Kinne, Oldendorf/Luhe     

Diurnal variation in chlorophyll fluorescence of Thalassia testudinum seedlings in response to controlled salinity and light conditions

Diurnal variability in chlorophyll fluorescence caused by dynamic irradiance conditions is an important issue when using pulse amplitude modulation fluorometry to measure physiological conditions of plants at the landscape scale. We examined the use of slopes and y-intercepts of diurnal effective photochemical efficiency of photosystem II (PSII) (ΔF/F _m′) versus photosynthetically active radiation (PAR) regressions in addition to direct measurements of maximum photochemical efficiencies of PSII (F _v/F _m) values to assess physiological status of Thalassia testudinum seedlings in a controlled mesocosm study. Seedlings were exposed to two light treatments (full sun and 50–70 % light reduction) and three salinity treatments (20, 35, and 50). Measurements were taken at 0600, 0900, 1200, 1500, 1800, and 2100 hours in order to assess the diurnal variation in photochemical efficiency of PSII and PAR, with measurements at 2100 providing F _v/F _m. Results indicated significant effects of light and salinity on regression y-intercepts and measured F _v/F _m values. Shaded seedlings had higher values for both parameters, suggesting low-light acclimation. The highest salinity treatment resulted in significant reductions for both parameters, suggesting stress. Stress was also indicated by significant reductions in both seedling leaf growth and mean differences between seedling leaves and media osmolalities in the hypersaline treatments (152.0 ± 26.4 vs. 630 ± 40.2 mmol kg^?1 for the control treatments). Slopes of ΔF/F _m′ versus PAR significantly differed with varying light treatments, with full sun seedlings exhibiting shallower slopes than shaded seedlings, indicating higher efficiency of dissipation of excess energy (photoprotection). These experimental results confirm field data suggesting that diurnal ΔF/F _m′ versus PAR regressions are responsive to changes in the physiological status of T. testudinum and that the y-intercepts of diurnal regressions may be used as a proxy for F _v/F _m.     

Fluorescence induction in the macroalgae Chondrus crispus (Rhodophyceae) and Ulva sp. (Chlorophyceae)

The induction of in vivo chlorophyll a (chl a) fluorescence (change in fluorescence intensity during a time-scale of ms to s) was measured to determine the potential of this technique for assessing the physiological condition of the macroalgae Chondrus crispus and Ulva sp. A gradient in variable fluorescence (P-F ₀ =peak minus initial fluorescence, a measure of Photosystem II activity) was found along the frond of C. crispus, the values increasing with distance from the thallus apex. No gradient was observed for Ulva sp. thalli. Nitrogen- or phosphorus-depleted Ulva sp. required a longer dark-conditioning period and had lower values of P-F ₀ than did controls. In contrast, no differences were found in P-F ₀ of N- or P-depleted C. crispus unless values were normalized to chl a. The irradiance history of C. crispus strongly influenced P-F ₀ , even after dark-conditioning: P-F ₀ declined by about 70% as the mean daily natural irradiance increased between 2 February and 14 March 1986; a negative correlation was observed between P-F ₀ and the photon flux density 1 d prior to the measurement; P-F ₀ remained elevated when C. crispus was grown under a low photon flux density; and P-F ₀ decreased in thalli within 5 d of transfer from growth under natural irradiance to an incubator with artificial irradiance. Changes in variable fluorescence at different growth irradiances of C. crispus may be due to adaptive changes in the relative absorption cross-section of this alga. The influence of irradiance history on Ulva sp. was minimal in comparison.Issued as NRCC No. 28730Part of this study was carried out while employed by Focal Marine Ltd., Bedford, Nova Scotia, Canada     

Age-specific responses to elevated salinity in the coastal marsh plant black needlerush (Juncus roemerianus Scheele) as determined through polyphasic chlorophyll a fluorescence transients (OJIP)

This study employed polyphasic chlorophyll a fluorescence transients (OJIP), a non-invasive marker of environmental stress in plants, to evaluate salt tolerance in three different Juncus roemerianus age classifications (6-, 24-, and 60-months). Following exposure to elevated salts (30 psu), the younger plants sustained growth, which was comparable to freshwater controls. While older (60-month) plants receiving only freshwater also grew over the 8-week study, the older salt-treated plants did not increase in size. Similarly, there were significant declines in variable chlorophyll a fluorescence parameters (F _v/F _m and F _v/F _o), electron transport flux per reaction center (ET_o/RC), and photosystem II performance index (PI_ABS) for 60-month J. roemerianus following salt treatment. These responses were not evident in the two younger salt-treated age classifications. Our results suggest that older J. roemerianus are less tolerant to rapid and sudden increases in salinity relative to younger plants and that this age-specific response may help explain observed discrepancies in salt tolerance in J. roemerianus.