ABSTRACT Benthic foraminifers that host algal symbionts are similar to corals in that they rely on their algal endosymbionts for their energy needs, calcify prolifically, and are sensitive to changes in environmental conditions. They are abundant in the benthos of coastal coral-reef areas and are found throughout the tropical and subtropical regions. Pulse Amplitude Modulated (PAM) chlorophyll fluorometry and chlorophyll a extraction techniques were used to quantify and compare the photosynthetic responses of the benthic foraminiferal, Archaias angulatus and their isolated endosymbionts, Chlamydomonas hedleyi, to short-term changes in temperature. Maximum quantum efficiency (Fv/Fm) and rapid light curves (RLCs), from which relative electron transport rates (rETR) of photosystem II (PSII) were derived, were investigated over a thermal range from 4.4° to 33.9 °C in three experiments that were 7 to 31 days in duration. Typical mean yields (Fv/Fm for healthy holobionts (symbionts in hospite) were 0.6 - 0.7, and for isolated symbionts 0.5 - 0.6. Chronic photoinhibition, indicated by significant decreases in Fv/Fm, occurred at temperatures above 31.0°C; there was minimal reduction in efficiency in cooler treatments. The trends between holobiont and symbionts were very similar in all of the photophysiological parameters measured [yield, photoefficiency (<α>), ETRmax and minimum saturating irradiance (Ek)] and supported the temperature range findings in terms of the tolerance of the specimens in the low temperatures up to 31.0 °C. For all photochemical measurements assessed, the holobiont values tended to be somewhat higher than those for the symbionts, with the exception of Ek, possibly indicating a tight coupling in the host-symbiont response during photosynthesis. Chlorophyll a (<μ>g/foram) was negatively correlated with temperature (r = -0.37, p < 0.001) in Experiments 1 and 2. However, in all 3 experiments, chlorophyll a was variable, suggesting a high degree of individual variability in A. angulatus and the ability to acclimate. Some differences observed among treatments may be related to differences in seasons when the specimens were collected and in length of time in culture prior to experiments. Holobiont median rETR light curve trends and photophysiological derived parameters recorded median Ek ranges of ~100-150 <μ>mol photons m-2 s-1, observed ETRmax light intensities of ~200 <μ>mol photons m-2 s-1 and photoinhibition, induced by increasing irradiance intensities, which occurred > 500 <μ>mol photons m-2 s-1. These light curve trends and derived parameters generally supported previous photosynthesis O2 and CO2 gas production studies of A. angulatus. The differences in responses associated with acclimation should be considered in design of future experimental studies. This was the first known physiological study of the viable temperature range and photobiology of A. angulatus using chlorophyll fluorometry methods. Though commonly found in Caribbean and Atlantic waters ranging from 14.0 - 31.0 °C, these results indicate a wider thermal-tolerance range for A. angulatus than was previously known. Keywords: Foraminifera, Chlamydomonas sp., PAM fluorometry, photosynthesis, algal symbiosis

Harmful algae blooms (HABs) have caused millions dollars in annual losses to the aquaculture industry, inhibited beach recreation, and have threatened marine and human health. HABs and red tides can develop suddenly and their frequency, geographic range, and intensity have increased over the past decade. A possible source for spreading and seeding new areas expanding the geographic range of HABs is ballast water. The process of ballast water discharge has been identified as a primary vector for the translocation of non-indigenous species (NIS) and invasive species. National and international efforts are currently underway to address the impact of NIS and invasive species. Policy is being developed detailing stringent rules to kill, remove, or otherwise inactive organisms in ballast water prior to or upon discharge. Currently, vendors are developing technologies to treat ballast water and U.S. and international facilities are testing these technologies to verify their efficacy. Ultraviolet (UV) radiation is commonly employed in ballast water treatment technologies. Previous studies have shown that UV light is effective for disinfecting drinking water, but the response of non-pathogenic and marine organisms is largely unknown. The purpose of this research was to measure the viability of the durable red-tide forming dinoflagellate, Lingulodinium polyedra following UV treatment. Two methods were used to measure the viability signal; manual epifluorescence microscopy with correlated viability stains and Pulse Amplitude Modulated (PAM) fluorometry to measure the physiological state of the organism following UV treatment. The number of cysts was also enumerated. The results showed that there was a significant decrease in the number of living L. polyedra cells following a UV treatment of more than 100 mWs cm-2. The results also have showed a significant increase in the number of L. polyedra cysts following UV treatment as low as 50 mWs cm-2.

Photo-oxidative stress is one of the key factors that can induce bleaching in reef organisms. With the decline of coral reefs and recurrent bleaching events, many studies have focused on understanding the mechanism behind this phenomenon. Two of the hypotheses that explain how the photosynthetic performance of the symbiont is affected and influences bleaching are: (1) disruption of the photosynthetic pathway by direct damage to the photosystem II (PSII), and (2) by inhibition of the Calvin-Benson cycle. In this dissertation I examine different aspects of photosynthetic performance in symbiont-bearing reef organisms and how this is influenced by symbiont loss and changes in photic stress as a result of different levels of irradiance modulated by time of the year (e.g., season) and depth; and take a closer look into primary productivity by symbionts with controlled laboratory experiments. Field experiments during 2012–2013 at Tennessee Reef, FL, assessed the photosynthetic performance of PSII in the diatom-bearing foraminifer, Amphistegina gibbosa, and the anthozoans: Palythoa cariabeorum, Siderastrea siderea, and Montastraea cavernosa. Data collected for the bleaching trends of A. gibbosa revealed that bleaching rates are higher in the summer months than in winter. Photochemical efficiencies of PSII in A. gibbosa, as measured with PAM fluorometry on the day of collection, were more variable in the shallow site (6 m) than in the deeper site (18 m). Also, photochemical efficiencies at the shallow site were lower during the summer months than during winter months. At the 18 m site, photochemical efficiencies did not exhibit a clear seasonal trend. Depth also had an effect on the measured photochemical efficiencies of the anthozoans. Photochemical efficiencies were lower and more variable in colonies at 6 m compared to colonies from 18 m. Although previous studies have reported seasonal effects on the photochemical efficiency of some coral colonies, that trend was not apparent in this study. Photoacclimation and productivity were assessed for A. gibbosa using rapid light curves (RLC) and photosynthesis vs. irradiance curves (P-E). Maximum relative electron transport rate (rETRmax) as described by RLCs was significantly different between A. gibbosa without visual signs of bleaching and those with severe bleaching. Individuals with partial bleaching had a rETRmax that was intermediate between the other two categories. The P-E curves showed a similar trend. In this case individuals that were non- or partly bleached had significantly higher photosynthesis maxima than those with severe bleaching. The onsets of photosynthesis and saturation irradiance were not significantly different among the categories of bleaching analyzed. Results from this dissertation suggest that A. gibbosa has the capability to detect and digest damaged symbionts, that the symbionts even in the deeper chambers react in a similar way to irradiance, but that in severe cases of bleaching the symbionts may not produce enough energy to sustain the requirements of the host, even in non-stressful conditions.

<p dir="ltr">Photosynthesis is the basis of primary productivity on Earth. Oxygenic photosynthesis utilizes the nearly inexhaustible energy of radiant solar light to fix atmospheric carbon dioxide into usable forms of chemical energy and produces dioxygen as a product. Central to this process are several large hetero-oligomeric protein complexes that comprise the photosynthetic electron transport chain. Photosystem II (PSII) initiates electron transport through the light-driven oxidation of water, in-turn relinquishing protons and oxygen. Through this reaction, electrons are used to form the reductant NADPH, while protons form a proton-motive gradient that is used to drive synthesis of ATP. As a result of this highly energetic reaction, PSII is often subject to oxidative photodamage due to the production of reactive oxygen species. Inevitably, accumulation of oxidative photodamage disrupts the catalytic activity of PSII, resulting in a loss of photosynthetic activity. To deal with the nearly constant incurred photodamage to PSII, oxygenic photoautotrophs undergo a disassembly and repair cycle that results in the complete turnover of the damaged D1 subunit of PSII. Due to its high tendency for damage, the D1 subunit has a half-life of under one hour in high light intensity. Despite our current understanding of photoinhibition and PSII repair, it is still unclear how D1 is replaced so rapidly in response to damaging conditions. Previous research has indicated a role for phosphorylation of PSII in D1 turnover, however the mechanism has not been totally resolved. In the first chapter of this thesis, our current understanding of PSII phosphorylation and oxidative damage is reviewed in the context of PSII repair. In the second chapter, the role of protein phosphorylation in the PSII repair cycle is investigated in the model organism <i>Arabidopsis</i>. Using several PSII phosphorylation mutants, we demonstrate that phosphorylation seems to mediate disassembly of large PSII supercomplexes and dimers into smaller subcomplexes. In the third chapter, the role of oxidative photodamage is investigated in mediating PSII disassembly. Here, we use several <i>in vitro</i> assays to demonstrate that photodamage is sufficient to induce the disassembly of smaller PSII subcomplexes. In the fourth chapter, a technique for determining the stoichiometry of photosynthetic complexes is examined, with implications for understanding PSII repair. Finally, in the fifth chapter, several conclusions and unanswered questions from this thesis are discussed.</p>

Snow algae are psychrophilic microorganisms, that inhabit snow fields in mountains and polar regions, which creates colored snow in good conditions. Most species belong to order Chlamydomonadales (Chlorophyta) with complicated life cycles, containing flagellates and immobile stages (cysts). Extreme environment of snow is characteristed by low temperature, problems with availability of water and lack of nutrition. Depending on location and phase of life cycle, snow algae have to deal with excess or lack of solar radiation, the important component of which is also dangerous UV radiation. The light conditions differ substantially from open location above the forest level or polar regions to forest habitats. In the first part of this work I compared reactions of photosynthetic apparatus of snow algae strains from forest and forestless habitats to different intensity of radiation, then I dealt with assessing any changes related to long-term cultivation in laboratory. Based on measurement of rapid light curves on PAM fluorometer I have determined some characteristics of photosynthetic apparatus of individual strains (parameters α and Ik), which indicate adaptation to low or high light intensity. For some strains, it was possible to compare the results obtained at a three-year interval. In the second part...

Macroalgal beds provide the ecological foundations for most shallow reef ecosystems in temperate environments. With distinctive canopies primarily of brown laminarian algae (northern hemisphere), or laminarian or fucalean algae (southern hemisphere), in many areas these habitats are at risk from human activity. Overexploitation, pollution, and other effects of coastal activities have resulted in significant habitat loss in coastal ecosystems, and human-induced climate change is now seen as a major threat to ecosystem health in marine systems. Understanding the impact of climate change is particularly important for habitat-forming ecosystem engineers like kelps, as these species form the basis of hierarchically organised communities and play a fundamental role in determining community structure and ecological processes. South eastern Australia has experienced increases in marine temperatures at nearly four times the global average, and there is now evidence that, in some locations, macroalgae communities are retreating in a manner consistent with ocean warming. Successful management of marine systems requires understanding ecosystem processes, particularly the patterns and magnitude of production. Macroalgal communities often show relatively low resistance to disturbance, yet rapid recovery once disturbances are removed, hence they are generally highly dynamic in response to environmental perturbations. As a result, macroalgae are likely to play an increasingly important role in buffering the short term/dynamic effects of climate change on temperate reef communities.Knowledge of the productivity of seaweed-dominated temperate reef systems is largely a synthesis from studies conducted over small spatial scales utilising a variety of methods that generally measure different characteristics of both individual seaweeds and collectively. As a result of the diversity of measurement methods, estimates of gross primary productivity (GPP), production potential, and macroalgal biomass for temperate reefs are numerous and variable. This can lead to challenges for ecologists attempting to amalgamate research findings to facilitate long-term, broad-scale perspectives or compare short-term research between spatially separated communities. However, to date there has been relatively little research to compare measurement approaches and quantify differences in productivity estimates across the different techniques.The present research provides a unique investigation into some of the techniques and methodology involved in measuring primary productivity in marine systems, particularly kelp forests, using the macroalgae Ecklonia radiata, Phyllospora comosa and Macrocystis pyrifera as study species. The work is based on both field and laboratory exploration of productivity measurements and associated parameters. In situ measurements of primary productivity (diel oxygen modelling, benthic oxygen exchange chambers) or PSII electron transport (PAM fluorometry) are compared, and the possibility of using acoustics as a means of quantifying oxygen production at large scales is explored, as has already been applied in seagrass beds. This thesis also provides an in depth investigation of the effect of variability in sampling methodology with regards to interpretation of PAM fluorometry-derived parameters. Chapter 2 investigates the acoustic properties of Ecklonia radiata. The density, sound speed and resulting adabiatic compressibility of E. radiata tissue were investigated in the laboratory. Four methods were developed and trialled to determine the intrinsic sound speed of Ecklonia radiata tissue based on measurement of the time of flight of an ultrasonic pulse, while compressibility was calculated from density measurements. The results show that Ecklonia radiata sound speed and density are higher, and compressibility lower, than that of seawater. Properties varied according to size and tissue type and the variation likely reflected differences in cell type, packing and structure as well as the concentrations of alginates and other carbohydrates. These are important considerations for acoustic propagation and the results provide valuable inputs for future acoustic work. Chapter 3 focuses on the acoustic modelling of different scenarios of primary production in a shallow water rocky reef environment of Fortescue Bay (Canoe Bay), Tasmania, where E. radiata dominates the canopy. In February 2012, the environment was continuously probed by acoustic signal transmission and monitored by a comprehensive set of oceanographic sensors with the aim to assess the potential for acoustics to quantify excess oxygen production in bubble form. Ray-theory acoustic modelling results indicate that ecologically-significant void fractions of oxygen in the canopy layer from production would be clearly seen in diel variation of propagation features such as the energy decay rate of the medium impulse response. The model can then be used to invert empirical data for retrieving void fraction. However, comparative analysis of part of FORTES 12 data and model suggests that no large excess of bubbles was produced by photosynthesis under the present environmental conditions, in contrast to earlier observations made in seagrasses. As a result, the use of acoustics as a means of measuring primary productivity in kelp could not be further explored during the course of this research.Chapter 4 provides a unique comparison of the estimates of photosynthetic O2 production rates in an Ecklonia radiata dominated community using three different measurement methods: diel oxygen GPP models, benthic oxygen exchange chambers, and electron transport rate from PAM fluorometry which is usually interpreted as a measure of production potential. All three methods were run concurrently in situ in Fortescue Bay, Tasmania. The first diel oxygen model was fitted to in situ measures of dissolved oxygen (DO) in the environment and demonstrated a good fit, however, a consequence of this approach is that large variation in oxygen production was predicted at low PAR levels. A second model was created which utilised an explicit relationship between DO production and PAR, but it didn’t represent DO at the surface as well as the first model. Importantly, the two models indicate similar daily production rates of the seaweed bed (all species combined) that are ~ 2 times that predicted for the kelp alone based on incubations in the benthic chambers and scaling for the average size of adult kelp sporophytes and their population density. Oxygen evolution from incubation of sporophytes in benthic chambers and PAM fluorometry derived electron transport rates showed similar patterns, but the results indicate that the latter method may overestimate potential photosynthesis. The results suggest that diel oxygen modelling, benthic oxygen exchange chambers and PAM fluorescence can all provide good indications of productivity in shallow water marine environments. However, care must be taken in interpretation of results as each method differs in the type of productivity estimates it provides. As a direct measure of total seaweed production per unit area of reef, estimates from models based on empirical measures of environmental DO have much to recommend them.Chapter 5 details a final analysis investigating the effects of diurnal, seasonal and latitudinal variability in ambient light on PAM-derived parameters, as well as possible effects associated with depth, within- and between-alga variation in PSII performance, and latitudinal effects unrelated to the light climate. This research was based on field measurements undertaken in Tasmania, Western Australia and New South Wales, Australia in both summer and winter during 2012 and 2013, focussing on Ecklonia radiata, Macrocystis pyrifera and Phyllospora comosa. Photosynthetic characteristics of all species were highly dependent on the time of day, depth, latitude/region, season, and part of the thallus from which measurements were taken. Patterns dependent on time-of-day, depth and thalli placement varied with season and/or geographic region, and the nature of these patterns varied between species. It is clear from this work that efforts to standardise approaches to taking measurements of seaweeds using PAM fluorometry will be essential if measurements are to be compared meaningfully across studies.The key findings of this thesis are: (1) a first determination of the acoustic properties of E. radiata tissue which enable the development of scattering models to interpret scientific echosounder data collected in kelp beds; (2) a Gaussian beam/finite element beam code (Bellhop) with detailed environmental input and a huge number of beams can predict the acoustic character of a shallow water rocky reef and bubble layers with low-frequency effective sound speed; (3) the model allows prediction of the acoustic energy decay rates due to various scenarios of ecologically-relevant photosynthetic O2 production rates; (4) day vs night acoustic measurement and model data comparisons challenge void fraction predictions made from well established measurements and methods; (5) diel oxygen modelling, benthic oxygen exchange chambers and PAM fluorescence can all provide good indications of productivity, however, understanding the limitations of each method is essential when interpreting the results as the measurements they provide are not directly comparable; and (6) applying a consistent sampling methodology is a key consideration when planning research utilising PAM fluorometry as diurnal, seasonal, and latitudinal variability, as well as effects associated with depth and within- and between-alga variation in PSII performance will have significant impact on PAM-derived parameters. The results of this work give valuable insight into the advantages and disadvantages involved with several main techniques currently utilised to measure production of macroalgal/seagrass beds, and the challenges faced by ecologists attempting to interpret results and compare research between methods and across studies. Last but not least, this study provides important and relevant information on the potential use of acoustics as a future means of determining productivity of benthic habitat on large scales in marine environments. The work presented herein will assist in both development and interpretation of future study of productivity in marine systems. / Doctorat en Sciences de l'ingénieur et technologie / info:eu-repo/semantics/nonPublished