Paul G. Falkowski

Using a high sensitivity differential scanning calorimeter in isothermal mode, we directly measured heat production in eukaryotic protists from 5 phyla spanning over 5 orders of magnitude in carbon biomass and 8 orders of magnitude in cell volume. Our results reveal that metabolic heat production normalized to cell mass is virtually constant in these organisms, with a median of 0.037 pW pg C ⁻¹ (95% confidence interval = 0.022–0.061 pW pg C ⁻¹ ) at 5 °C. Contrary to allometric models, the relationship between heat production and cell carbon content or surface area is isometric (scaling exponents, 1.056 and 1.057, respectively). That heat production per unit cell surface area is constant suggests that heat flux through the cell surface is effectively instantaneous, and hence that cells are isothermal with their environment. The results further suggest that allometric models of metabolism based on metazoans are not applicable to protists, and that the underlying metabolic processes in the latter polyphyletic group are highly constrained by evolutionary selection. We propose that the evolutionary constraint leading to a universally constant heat production in single-celled eukaryotes is related to cytoplasmic packaging of organelles and surface area to volume relationships controlling diffusion of resources to these organelles.

Carbon uptake by marine phytoplankton, and its export as organic matter to the ocean interior (i.e., the “biological pump”), lowers the partial pressure of carbon dioxide ( p CO ₂ ) in the upper ocean and facilitates the diffusive drawdown of atmospheric CO ₂ . Conversely, precipitation of calcium carbonate by marine planktonic calcifiers such as coccolithophorids increases p CO ₂ and promotes its outgassing (i.e., the “alkalinity pump”). Over the past ≈100 million years, these two carbon fluxes have been modulated by the relative abundance of diatoms and coccolithophores, resulting in biological feedback on atmospheric CO ₂ and Earth's climate; yet, the processes determining the relative distribution of these two phytoplankton taxa remain poorly understood. We analyzed phytoplankton community composition in the Atlantic Ocean and show that the distribution of diatoms and coccolithophorids is correlated with the nutricline depth, a proxy of nutrient supply to the upper mixed layer of the ocean. Using this analysis in conjunction with a coupled atmosphere–ocean intermediate complexity model, we predict a dramatic reduction in the nutrient supply to the euphotic layer in the coming century as a result of increased thermal stratification. Our findings indicate that, by altering phytoplankton community composition, this causal relationship may lead to a decreased efficiency of the biological pump in sequestering atmospheric CO ₂ , implying a positive feedback in the climate system. These results provide a mechanistic basis for understanding the connection between upper ocean dynamics, the calcium carbonate-to-organic C production ratio and atmospheric p CO ₂ variations on time scales ranging from seasonal cycles to geological transitions.

How did biological, geochemical, and geophysical processes produce an atmosphere that allowed complex animal life to evolve?

Superoxide dismutase (SOD) catalyzes the transformation of superoxide to molecular oxygen and hydrogen peroxide. Of the four known SOD isoforms, distinguished by their metal cofactor (iron, manganese [Mn], copper/zinc, nickel), MnSOD is the dominant form in the diatom Thalassiosira pseudonana. We cloned the MnSOD gene, sodA, using the expression vector pBAD, overexpressed the product in Escherichia coli, and purified the mature protein (TpMnSOD). This recombinant enzyme was used to generate a polyclonal antibody in rabbit that recognizes MnSOD in T. pseudonana. Based on quantitative immunoblots, we calculate that in vivo concentrations of TpMnSOD are approximately 0.9 amol cell(-1) using the recombinant protein as a standard. Immunogold staining indicates that TpMnSOD is localized in the chloroplasts, which is in contrast to most other eukaryotic algae (including chlorophytes and embryophytes) where MnSOD is localized exclusively in mitochondria. Based on the photosynthetic Mn complex in photosystem II, cellular Mn budgets cannot account for 50% to 80% of measured Mn within diatom cells. Our results reveal that chloroplastic MnSOD accounts for 10% to 20% of cellular Mn, depending on incident light intensity and cellular growth rate. Indeed, our analysis indicates that TpMnSOD accounts for 1.4% (+/-0.2%) of the total protein in the cell. The TpMnSOD has a rapid turnover rate with an apparent half-life of 6 to 8 h when grown under continuous light. TpMnSOD concentrations increase relative to chlorophyll, with an increase in incident light intensity to minimize photosynthetic oxidative stress. The employment of a Mn-based SOD, linked to photosynthetic stress in T. pseudonana, may contribute to the continued success of diatoms in the low iron regions of the modern ocean.

As oxygen built up in Earth's atmosphere during the Precambrian, organisms evolved more complex biochemical networks. This expansion made feasible oxygen dependent metabolisms that coopted parts of preexisting anaerobic ones.

On the basis of a carbon isotopic record of both marine carbonates and organic matter from the Triassic-Jurassic boundary to the present, we modeled oxygen concentrations over the past 205 million years. Our analysis indicates that atmospheric oxygen approximately doubled over this period, with relatively rapid increases in the early Jurassic and the Eocene. We suggest that the overall increase in oxygen, mediated by the formation of passive continental margins along the Atlantic Ocean during the opening phase of the current Wilson cycle, was a critical factor in the evolution, radiation, and subsequent increase in average size of placental mammals.

Using a bioinformatic approach, we analyzed the correspondence in genetic distance matrices between all possible pairwise combinations of 82 photosynthetic genes in 10 species of cyanobacteria. Our analysis reveals significant correlations between proteins linked in a conserved gene order and between structurally identified interacting protein scaffolds that coordinate the binding of cofactors involved in photosynthetic electron transport. Analyses of amino acid substitution rates suggest that the tempo of evolution of genes encoding core metabolic processes in the photosynthetic apparatus is highly constrained by protein-protein, protein-lipid, and protein-cofactor interactions (collectively called “protein interactions”). These interactions are critical for energy transduction, primary charge separation, and electron transport and effectively act as an internal selection pressure governing the conservation of clusters of photosynthetic genes in oxygenic prokaryotic photoautotrophs. Consequently, although several proteins within the photosynthetic apparatus are biophysically and physiologically inefficient, selection has not significantly altered the genes encoding these essential proteins over billions of years of evolution. In effect, these core proteins have become “frozen metabolic accidents.”

We identify four novel DNA-binding complexes in the nuclear-encoded Lhcb1 promoter of the chlorophyte alga Dunaliella tertiolecta that are regulated by photosynthetic pathways in the plastid. The binding activities of three of the complexes were positively correlated with time-dependent changes in Lhcb1 transcript abundance, implicating their roles as transcriptional enhancers in a retrograde signal transduction pathway. Using a combination of inhibitors, uncouplers, and antimycin A, and by following the kinetic pattern of gene regulation, we infer two different sensors in the signal transduction pathway. On short time scales of 0.5 to about 4 h, the transthylakoid membrane potential appears to be a critical determinant of gene expression, whereas on time scales of 8 h or longer, the redox state of the plastoquinone pool becomes increasingly more important. The differentiation of these two types of signals was observed in parallel effects on gene transcription and on the patterns of DNA-binding activities in the Lhcb1 promoter. These signals appear to be transduced at the nuclear level via a coordinated ensemble of DNA-binding complexes located between −367 and −188 bp from the start codon of the gene. The regulation of these elements allows the cell to up- or down-regulate the expression on Lhcb1 in response to changes in irradiance.

Over the past three decades, massive bleaching events of zooxanthellate corals have been documented across the range of global distribution. Although the phenomenon is correlated with relatively small increases in sea-surface temperature and enhanced light intensity, the underlying physiological mechanism remains unknown. In this article we demonstrate that thylakoid membrane lipid composition is a key determinate of thermal-stress sensitivity in symbiotic algae of cnidarians. Analyses of thylakoid membranes reveal that the critical threshold temperature separating thermally tolerant from sensitive species of zooxanthellae is determined by the saturation of the lipids. The lipid composition is potentially diagnostic of the differential nature of thermally induced bleaching found in scleractinian corals. Measurements of variable chlorophyll fluorescence kinetic transients indicate that thermally damaged membranes are energetically uncoupled but remain capable of splitting water. Consequently, a fraction of the photosynthetically produced oxygen is reduced by photosystem I through the Mehler reaction to form reactive oxygen species, which rapidly accumulate at high irradiance levels and trigger death and expulsion of the endosymbiotic algae. Differential sensitivity to thermal stress among the various species of Symbiodinium seems to be distributed across all clades. A clocked molecular phylogenetic analysis suggests that the evolutionary history of symbiotic algae in cnidarians selected for a reduced tolerance to elevated temperatures in the latter portion of the Cenozoic.