Mark E. Warner

EFFECTS OF INCREASED TEMPERATURE AND CO2 ON PHOTOSYNTHESIS, GROWTH, AND ELEMENTAL RATIOS IN MARINE SYNECHOCOCCUS AND PROCHLOROCOCCUS (CYANOBACTERIA)1

Little is known about the combined impacts of future CO2 and temperature increases on the growth and physiology of marine picocyanobacteria. We incubated Synechococcus and Prochlorococcus under present‐day (380 ppm) or predicted year‐2100 CO2 levels (750 ppm), and under normal versus elevated temperatures (+4°C) in semicontinuous cultures. Increased temperature stimulated the cell division rates of Synechococcus but not Prochlorococcus. Doubled CO2 combined with elevated temperature increased maximum chl a–normalized photosynthetic rates of Synechococcus four times relative to controls. Temperature also altered other photosynthetic parameters (α, Φmax, Ek, and ) in Synechococcus, but these changes were not observed for Prochlorococcus. Both increased CO2 and temperature raised the phycobilin and chl a content of Synechococcus, while only elevated temperature increased divinyl chl a in Prochlorococcus. Cellular carbon (C) and nitrogen (N) quotas, but not phosphorus (P) quotas, increased with elevated CO2 in Synechococcus, leading to ∼20% higher C:P and N:P ratios. In contrast, Prochlorococcus elemental composition remained unaffected by CO2, but cell volume and elemental quotas doubled with increasing temperature while maintaining constant stoichiometry. Synechococcus showed a much greater response to CO2 and temperature increases for most parameters measured, compared with Prochlorococcus. Our results suggest that global change could influence the dominance of Synechococcus and Prochlorococcus ecotypes, with likely effects on oligotrophic food‐web structure. However, individual picocyanobacteria strains may respond quite differently to future CO2 and temperature increases, and caution is needed when generalizing their responses to global change in the ocean.

Interactive effects of increased pCO 2 , temperature and irradiance on the marine coccolithophore Emiliania huxleyi (Prymnesiophyceae)

We examined the effects of increased temperature, pCO2, and irradiance on a calcifying strain of the marine coccolithophore Emiliania huxleyi in semi-continuous laboratory cultures. Emiliania huxleyi CCMP 371 was cultured in four temperature and pCO2 treatments at both low and high irradiance (50 and 400 µmol photons m−2 s−1): (i) 20°C and 375 ppm CO2 (ambient control); (ii) 20°C and 750 ppm CO2 (high pCO2); (iii) 24°C and 375 ppm CO2 (high temperature); and (iv) 24°C and 750 ppm CO2 (‘greenhouse’). The growth of E. huxleyi was greatly accelerated by elevated temperature at low irradiance. Photosynthesis was significantly promoted by increases in both pCO2 and temperature at both irradiances. Higher cellular C/P ratios were found in the higher CO2 treatments at high irradiance, indicating a reduced requirement for P. The PIC/POC (particulate inorganic to organic carbon) ratio remained constant at low light, regardless of CO2 or temperature conditions. However, both the cellular PIC content and PIC/POC ratio were greatly decreased by elevated irradiance, and were further decreased by increased pCO2 only at high light, indicating a combined effect of CO2 and light on calcification. These results suggest that future trends of CO2 enrichment, sea-surface warming and exposure to higher mean irradiances from intensified stratification will have a large influence on the growth of Emiliania huxleyi, and potentially on the PIC/POC ‘rain ratio’. Our study demonstrates that it is possible to obtain a more complete picture of global change impacts on marine phytoplankton by designing experiments that consider multiple global change variables and their mutual interactions.

DIFFERENTIAL IMPACTS OF PHOTOACCLIMATION AND THERMAL STRESS ON THE PHOTOBIOLOGY OF FOUR DIFFERENT PHYLOTYPES OF SYMBIODINIUM (PYRRHOPHYTA)1

The capacity for photoacclimation to light at 100 or 600 μmol photons·m−2·s−1 and the subsequent response to thermal stress was examined in four genetically distinct cultures of symbiotic dinoflagellates in the genus Symbiodinium with the ITS2 designations A1, A1.1, B1, and F2. While all algal types showed typical signs of photoacclimation to high light via a reduction in chl a, there was a differential response in cellular growth, photosystem II (PSII) activity, and the chl a‐specific absorption coefficient between cultures. When maintained at 32°C for up to 10 days, significant variation in the susceptibility to thermal stress was observed in the rate of loss in PSII activity and electron transport, PSII reaction center degradation, and cellular growth. The order of thermal tolerance did not change between the two light levels. However, as expected, loss in photosynthetic function was exacerbated in the thermally sensitive phylotypes (B1 and A1.1) when acclimated to the higher light intensity. There was no consistent relationship between thermal tolerance and changes in light energy dissipation via non‐photochemical pathways. Phylotypes F2 and A1 showed a high degree of thermal tolerance, yet the cellular responses to light and temperature were markedly different between these algae. The F2 isolate showed the greatest capacity for photoacclimation and growth at high light and temperature, while the A1 isolate appeared to adjust to thermal stress by a slight decline in PSII activity and a significant decline in growth, possibly at the expense of increased photosystem and cellular repair rates.

Photosynthesis and production of hydrogen peroxide by Symbiodinium (Pyrrhophyta) phylotypes with different thermal tolerances

Occurrences whereby cnidaria lose their symbiotic dinoflagellate microalgae (Symbiodinium spp.) are increasing in frequency and intensity. These so‐called bleaching events are most often related to an increase in water temperature, which is thought to limit certain Symbiodinium phylotypes from effectively dissipating absorbed excitation energy that is otherwise used for photochemistry. Here, we examined photosynthetic characteristics and hydrogen peroxide (H2O2) production, a possible signal involved in bleaching, from two Symbiodinium types (a thermally “tolerant” A1 and “sensitive” B1) representative of cnidaria–Symbiodinium symbioses of reef‐building Caribbean corals. Under steady‐state growth at 26°C, a higher efficiency of PSII photochemistry, rate of electron turnover, and rate of O2 production were observed for type A1 than for B1. The two types responded very differently to a period of elevated temperature (32°C): type A1 increased light‐driven O2 consumption but not the amount of H2O2 produced; in contrast, type B1 increased the amount of H2O2 produced without an increase in light‐driven O2 consumption. Therefore, our results are consistent with previous suggestions that the thermal tolerance of Symbiodinium is related to adaptive constraints associated with photosynthesis and that sensitive phylotypes are more prone to H2O2 production. Understanding these adaptive differences in the genus Symbiodinium will be crucial if we are to interpret the response of symbiotic associations, including reef‐building corals, to environmental change.

The Relative Significance of Host–Habitat, Depth, and Geography on the Ecology, Endemism, and Speciation of Coral Endosymbionts in the Genus Symbiodinium

Dinoflagellates in the genus Symbiodinium are among the most abundant and important group of eukaryotic microbes found in coral reef ecosystems. Recent analyses conducted on various host cnidarians indicated that Symbiodinium assemblages in the Caribbean Sea are genetically and ecologically diverse. In order to further characterize this diversity and identify processes important to its origins, samples from six orders of Cnidaria comprising 45 genera were collected from reef habitats around Barbados (eastern Caribbean) and from the Mesoamerican barrier reef off the coast of Belize (western Caribbean). Fingerprinting of the ribosomal internal transcribed spacer 2 identified 62 genetically different Symbiodinium. Additional analyses of clade B Symbiodinium using microsatellite flanker sequences unequivocally characterized divergent lineages, or “species,” within what was previously thought to be a single entity (B1 or B184). In contrast to the Indo-Pacific where host-generalist symbionts dominate many coral communities, partner specificity in the Caribbean is relatively high and is influenced little by the host’s apparent mode of symbiont acquisition. Habitat depth (ambient light) and geographic isolation appeared to influence the bathymetric zonation and regional distribution for most of the Symbiodinium spp. characterized. Approximately 80% of Symbiodinium types were endemic to either the eastern or western Caribbean and 40–50% were distributed to compatible hosts living in shallow, high-irradiance, or deep, low-irradiance environments. These ecologic, geographic, and phylogenetic patterns indicate that most of the present Symbiodinium diversity probably originated from adaptive radiations driven by ecological specialization in separate Caribbean regions during the Pliocene and Pleistocene periods.

Photobiology of symbiodinium revisited: bio-physical and bio-optical signatures

Light is often the most abundant resource within the nutrient-poor waters surrounding coral reefs. Consequently, zooxanthellae (Symbiodinium spp.) must continually photoacclimate to optimise productivity and ensure coral success. In situ coral photobiology is becoming dominated by routine assessments using state-of-the-art non-invasive bio-optical or chlorophyll a fluorescence (bio-physical) techniques. Multiple genetic types of Symbiodinium are now known to exist; however, little focus has been given as to how these types differ in terms of characteristics that are observable using these techniques. Therefore, this investigation aimed to revisit and expand upon a pivotal study by Iglesias-Prieto and Trench (1994) by comparing the photoacclimation characteristics of different Symbiodinium types based on their bio-physical (chlorophyll a fluorescence, reaction centre counts) and bio-optical (optical absorption, pigment concentrations) ‘signatures’. Signatures described here are unique to Symbiodinium type and describe phenotypic responses to set conditions, and hence are not suitable to describe taxonomic structure of in hospiteSymbiodinium communities. In this study, eight Symbiodinium types from clades and sub-clades (A–B, F) were grown under two PFDs (Photon Flux Density) and examined. The photoacclimation response by Symbiodinium was highly variable between algal types for all bio-physical and for many bio-optical measurements; however, a general preference to modifying reaction centre content over effective antennae-absorption was observed. Certain bio-optically derived patterns, such as light absorption, were independent of algal type and, when considered per photosystem, were matched by reaction centre stoichiometry. Only by better understanding genotypic and phenotypic variability between Symbiodinium types can future studies account for the relative taxonomic and physiological contribution by Symbiodinium to coral acclimation.

Coral Energy Reserves and Calcification in a High-CO2 World at Two Temperatures

Rising atmospheric CO2 concentrations threaten coral reefs globally by causing ocean acidification (OA) and warming. Yet, the combined effects of elevated pCO2 and temperature on coral physiology and resilience remain poorly understood. While coral calcification and energy reserves are important health indicators, no studies to date have measured energy reserve pools (i.e., lipid, protein, and carbohydrate) together with calcification under OA conditions under different temperature scenarios. Four coral species, Acropora millepora, Montipora monasteriata, Pocillopora damicornis, Turbinaria reniformis, were reared under a total of six conditions for 3.5 weeks, representing three pCO2 levels (382, 607, 741 µatm), and two temperature regimes (26.5, 29.0°C) within each pCO2 level. After one month under experimental conditions, only A. millepora decreased calcification (−53%) in response to seawater pCO2 expected by the end of this century, whereas the other three species maintained calcification rates even when both pCO2 and temperature were elevated. Coral energy reserves showed mixed responses to elevated pCO2 and temperature, and were either unaffected or displayed nonlinear responses with both the lowest and highest concentrations often observed at the mid-pCO2 level of 607 µatm. Biweekly feeding may have helped corals maintain calcification rates and energy reserves under these conditions. Temperature often modulated the response of many aspects of coral physiology to OA, and both mitigated and worsened pCO2 effects. This demonstrates for the first time that coral energy reserves are generally not metabolized to sustain calcification under OA, which has important implications for coral health and bleaching resilience in a high-CO2 world. Overall, these findings suggest that some corals could be more resistant to simultaneously warming and acidifying oceans than previously expected.

A Connection between Colony Biomass and Death in Caribbean Reef-Building Corals

Increased sea-surface temperatures linked to warming climate threaten coral reef ecosystems globally. To better understand how corals and their endosymbiotic dinoflagellates (Symbiodinium spp.) respond to environmental change, tissue biomass and Symbiodinium density of seven coral species were measured on various reefs approximately every four months for up to thirteen years in the Upper Florida Keys, United States (1994–2007), eleven years in the Exuma Cays, Bahamas (1995–2006), and four years in Puerto Morelos, Mexico (2003–2007). For six out of seven coral species, tissue biomass correlated with Symbiodinium density. Within a particular coral species, tissue biomasses and Symbiodinium densities varied regionally according to the following trends: Mexico≥Florida Keys≥Bahamas. Average tissue biomasses and symbiont cell densities were generally higher in shallow habitats (1–4 m) compared to deeper-dwelling conspecifics (12–15 m). Most colonies that were sampled displayed seasonal fluctuations in biomass and endosymbiont density related to annual temperature variations. During the bleaching episodes of 1998 and 2005, five out of seven species that were exposed to unusually high temperatures exhibited significant decreases in symbiotic algae that, in certain cases, preceded further decreases in tissue biomass. Following bleaching, Montastraea spp. colonies with low relative biomass levels died, whereas colonies with higher biomass levels survived. Bleaching- or disease-associated mortality was also observed in Acropora cervicornis colonies; compared to A. palmata, all A. cervicornis colonies experienced low biomass values. Such patterns suggest that Montastraea spp. and possibly other coral species with relatively low biomass experience increased susceptibility to death following bleaching or other stressors than do conspecifics with higher tissue biomass levels.

CONCENTRATIONS OF DIMETHYLSULFONIOPROPIONATE AND DIMETHYL SULFIDE ARE STRAIN‐SPECIFIC IN SYMBIOTIC DINOFLAGELLATES (SYMBIODINIUM SP., DINOPHYCEAE)1

Dimethyl sulfide (DMS) and dimethylsulfoniopropionate (DMSP) are sulfur compounds that may function as antioxidants in algae. Symbiotic dinoflagellates of the genus Symbiodinium show strain‐specific differences in their susceptibility to temperature‐induced oxidative stress and have been shown to contain high concentrations of DMSP. We investigated continuous cultures of four strains from distinct phylotypes (A1, A13, A2, and B1) that can be characterized by differential thermal tolerances. We hypothesized that strains with high thermal tolerance have higher concentrations of DMSP and DMS in comparison to strains with low thermal tolerance. DMSP concentrations were strain‐specific with highest concentrations occurring in A1 (225 ± 3.5 mmol · L−1  cell volume [CV]) and lowest in A2 (158 ± 3.8 mmol · L−1 CV). Both strains have high thermal tolerance. Strains with low thermal tolerance (A13 and B1) showed DMSP concentrations in between these extremes (194 ± 19.0 and 160 ± 6.1 mmol · L−1  CV, respectively). DMS data further confirmed this general pattern with high DMS concentrations in A1 and A13 (4.1 ± 1.22 and 2.1 ± 0.37 mmol · L−1 CV, respectively) and low DMS concentrations in A2 and B1 (0.3 ± 0.06 and 0.5 ± 0.22 mmol · L−1 CV, respectively). Hence, the strain‐specific differences in DMSP and DMS concentrations did not match the different abilities of the four phylotypes to withstand thermal stress. Future work should quantify the possible dynamics in DMSP and DMS concentrations during periods of high oxidative stress in Symbiodinium sp. and address the role of these antioxidants in zooxanthellate cnidarians.

Spring “bleaching” among Pocillopora in the Sea of Cortez, Eastern Pacific

A mild bleaching event was observed among Pocillopora spp. in the southern Gulf of California in the spring of 2006. Uniform bleaching occurred in numerous colonies on the upper portions of their branches. Most (∼90%) colonies that exhibited bleaching contained a species of endosymbiotic dinoflagellate, Symbiodinium C1b-c, which differed from the Symbiodinium D1 found inhabiting most unbleached colonies. Analysis of chlorophyll fluorescence, indicated a decline in photosystem II photochemical activity, especially among colonies populated with C1b-c. By early August, most affected colonies had recovered their normal pigmentation and fluorescence values were once again high for all colonies. No mortality was observed among tagged bleached colonies nor did symbiont species composition change during recovery. This unusual episode of bleaching did not appear to be a response to thermal stress, but may have been triggered by high levels of solar radiation during a period of unseasonally high water clarity in the early spring.