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In reef-building corals (order Scleractinia) and giant clams (phylum Molluca), V-type H+-ATPase (VHA) in host cells is part of a carbon concentrating mechanism (CCM) that regulates photosynthetic rates of their symbiotic algae. Here, we show that VHA plays a similar role in the sea anemone Anemonia majano, a member of the order Actinaria and sister group to the Scleractinia, which in contrast to their colonial calcifying coral relatives is a solitary, soft-bodied taxa. Western blotting and immunofluorescence revealed that VHA was abundantly present in the host-derived symbiosome membrane surrounding the photosymbionts. Pharmacological inhibition of VHA activity in individual anemones resulted in an approximately 80% decrease of photosynthetic O2 production. These results extend the presence of a host-controlled VHA-dependent CCM to non-calcifying cnidarians of the order Actiniaria, suggesting it is widespread among photosymbiosis between aquatic invertebrates and Symbiodiniaceae algae.

In reef-building corals (order Scleractinia) and giant clams (phylum Molluca), V-type H+-ATPase (VHA) in host cells is part of a carbon concentrating mechanism (CCM) that regulates photosynthetic rates of their symbiotic algae. Here, we show that VHA plays a similar role in the sea anemone Anemonia majano, a member of the order Actinaria and sister group to the Scleractinia, which in contrast to their colonial calcifying coral relatives is a solitary, soft-bodied taxa. Western blotting and immunofluorescence revealed that VHA was abundantly present in the host-derived symbiosome membrane surrounding the photosymbionts. Pharmacological inhibition of VHA activity in individual anemones resulted in an approximately 80% decrease of photosynthetic O2 production. These results extend the presence of a host-controlled VHA-dependent CCM to non-calcifying cnidarians of the order Actiniaria, suggesting it is widespread among photosymbiosis between aquatic invertebrates and Symbiodiniaceae algae.

A field dataset detailing the abundance and carbon use strategies (obtained with C13 isotopes) of marine macroalgae within 3 wave-exposed and 4 wave-sheltered coastal kelp forests in Otago, New Zealand, along with site information such as temperature, light, nutrients of each site. Additionally, a dataset of the results from a laboratory experiment measuring the photosynthetic response of Xiphophora gladiata and Hymenena palmata to increasing DIC concentrations and under high or low water motion and high (8.1) or low (7.6) pH.

We explored the metabolic pathways in two industrially relevant marine microorganisms to understand their core metabolic capabilities. It is necessary to track how an organism distributes organic building blocks throughout its metabolic pathways so that we can devise strategies to alter its metabolism and reroute substantial metabolic flux towards target compound(s). Though we cannot measure intracellular metabolic fluxes directly, we can retro-biosynthetically calculate them by supplying substrates labeled with non-radioactive isotopes to an organism. We then measure the resulting isotope labeling patterns of metabolites and calculate the fluxes that produced them. We addressed three goals with our research, (i) resolving questions surrounding organic carbon metabolism in the diatom Phaeodactylum tricornutum (P. tricornutum), (ii) identifying reactions in a putative photosynthetic carbon concentrating mechanism in P. tricornutum and (iii) mapping central carbon metabolism of the cellulolytic aerobe Saccharophagus degradans (S. degradans). Towards goal (i) we show that P. tricornutum predominantly consumes glucose, as opposed to atmospheric CO2, under mixotrophic conditions using the Entner-Doudoroff (ED) glycolytic pathway instead of the more common Embden-Meyerhof-Parnas pathway (EMP). We utilized metabolic flux analysis (MFA) to discover that acetate is metabolized for energy production instead of for biomass formation during mixotrophic growth on CO2 and acetate. Finally, we developed a method for measuring isotopic labeling in polyunsaturated fatty acids via gas chromatography-mass spectrometry (GC-MS), and demonstrated its utility in resolving outstanding questions about glucose metabolism by P. tricornutum. Towards goal (ii) we utilized isotope labeling and gene silencing in combination to identify pyruvate carboxylase as a key enzyme in a C4 carbon concentrating mechanism in P. tricornutum, while also ruling out phosphoenolpyruvate carboxylase as a key enzym

Marine diatoms play a critical role in the global carbon cycle, where they are responsible for 20% of all primary production. RubisCO, the rate limiting enzyme in carbon fixation, has a half-saturation constant many times higher than oceanic CO2 concentrations. In order to overcome this limitation, diatom species have evolved a diverse array of highly efficient carbon concentrating mechanisms. Increasing our understanding of these mechanisms provides a foundation to improve genetic engineering of these organisms for biofuel production and increased carbon sequestration, as well as a basis to potentially improve the efficiency of photosynthesis in terrestrial plants.In this study, the carbon concentrating mechanism of the diatom species Phaeodactylum tricornutum is investigated through kinetic modeling of potential pathways and analysis of differential gene expression of CCM-related genes. An existing kinetic model is rebuilt and expanded to explore proposed carbon-concentrating mechanisms and provide predictive values of carbon fluxes through the system at varying external conditions. The feasibility of each of three potential mechanisms is evaluated. Through gene expression analysis, potential major regulators of the carbon concentrating mechanism are identified.This investigation finds that the presence of bicarbonate transporters on the plasmalemma, CER, and chloroplast membranes is needed for the CCM to achieve expected carbon uptake and photosynthetic fluxes, but a bicarbonate transporter on the PPC membrane is not necessary. Additionally, the down-regulation of a chloroplast-membrane targeted bicarbonate transporter at sub-atmospheric CO2 indicates that a secondary CCM pathway takes over under low carbon stress.

Marine diatoms play a critical role in the global carbon cycle, where they are responsible for 20% of all primary production. RubisCO, the rate limiting enzyme in carbon fixation, has a half-saturation constant many times higher than oceanic CO2 concentrations. In order to overcome this limitation, diatom species have evolved a diverse array of highly efficient carbon concentrating mechanisms. Increasing our understanding of these mechanisms provides a foundation to improve genetic engineering of these organisms for biofuel production and increased carbon sequestration, as well as a basis to potentially improve the efficiency of photosynthesis in terrestrial plants.In this study, the carbon concentrating mechanism of the diatom species Phaeodactylum tricornutum is investigated through kinetic modeling of potential pathways and analysis of differential gene expression of CCM-related genes. An existing kinetic model is rebuilt and expanded to explore proposed carbon-concentrating mechanisms and provide predictive values of carbon fluxes through the system at varying external conditions. The feasibility of each of three potential mechanisms is evaluated. Through gene expression analysis, potential major regulators of the carbon concentrating mechanism are identified.This investigation finds that the presence of bicarbonate transporters on the plasmalemma, CER, and chloroplast membranes is needed for the CCM to achieve expected carbon uptake and photosynthetic fluxes, but a bicarbonate transporter on the PPC membrane is not necessary. Additionally, the down-regulation of a chloroplast-membrane targeted bicarbonate transporter at sub-atmospheric CO2 indicates that a secondary CCM pathway takes over under low carbon stress.

Symbiotic dinoflagellate algae residing inside coral tissues supply the host with the majority of their energy requirements through the translocation of photosynthetically fixed carbon. The algae, in turn, rely on the host for the supply of inorganic carbon. Carbon must be concentrated as CO2 in order for photosynthesis to proceed, and here we show that the coral host plays an active role in this process. The host-derived symbiosome membrane surrounding the algae abundantly expresses vacuolar H(+)-ATPase (VHA), which acidifies the symbiosome space down to pH ∼ 4. Inhibition of VHA results in a significant decrease in average H(+) activity in the symbiosome of up to 75% and a significant reduction in O2 production rate, a measure of photosynthetic activity. These results suggest that host VHA is part of a previously unidentified carbon concentrating mechanism for algal photosynthesis and provide mechanistic evidence that coral host cells can actively modulate the physiology of their symbionts.

Symbiotic dinoflagellate algae residing inside coral tissues supply the host with the majority of their energy requirements through the translocation of photosynthetically fixed carbon. The algae, in turn, rely on the host for the supply of inorganic carbon. Carbon must be concentrated as CO2 in order for photosynthesis to proceed, and here we show that the coral host plays an active role in this process. The host-derived symbiosome membrane surrounding the algae abundantly expresses vacuolar H(+)-ATPase (VHA), which acidifies the symbiosome space down to pH ∼ 4. Inhibition of VHA results in a significant decrease in average H(+) activity in the symbiosome of up to 75% and a significant reduction in O2 production rate, a measure of photosynthetic activity. These results suggest that host VHA is part of a previously unidentified carbon concentrating mechanism for algal photosynthesis and provide mechanistic evidence that coral host cells can actively modulate the physiology of their symbionts.

Photosynthesis is crucial for life but is a slow process because the CO2 concentration near the principal carbon-assimilation enzyme RuBisCO is extremely low. Very few plants and algae perform a carbon-concentrating mechanism (CCM) to overcome the insufficiency, which are classified into biophysical and biochemical (C4) mechanism. The enzyme CA catalyzes the reversible dehydration of HCO3- to CO2 in biophysical CCMs and its active site contains a Zn2+. In this study, we hypothesized that Zn2+ availability can impact CCMs and therefore investigated the effect of Zn2+ availability on photosynthetic metabolism in a unicellular marine diatom Phaeodactylum tricornutum. P. tricornutum has a sequenced genome and can conduct both biophysical and C4 CCMs. We observed that Zn2+ has a significant effect on cell growth rate but no significant interference on intracellular metabolism, suggesting no essential compensation of C4 CCMs for biophysical CCMs even at low CA activity anticipated at low Zn2+ concentration.

We have assessed the effect of vanadate as an inhibitor of plasma-membrane ATPase on photosynthesis and the ATP/ADP ratio in Chlamydomonas reinhardtii CW-92 (a mutant strain lacking a cell wall). This effect was compared in low-CO2-adapted cells grown in media bubbled with air containing 400 or 70 muL . L-1 CO2. Evidence is presented indicating that cells grown at 70 muL . L-1 CO2 have a higher rate of photosynthetic O2 evolution than cells grown at 400 muL . L-1 CO2, at limiting carbon concentrations. Extracellular and intracellular carbonic-anhydrase activities were, however, similar in cells grown in both of the low-carbon conditions. Vanadate inhibited, to a different extent, the HCO3--dependent O2 evolution in cells grown at 400 and 70 muL . L-1 CO2. At 400 muM vanadate, inhibition reached 70-75 % in cells grown at 400 muL . L-1 but only 50 % in those grown at 70 muL . L-1 CO2. The ATP/ADP ratios determined with and without vanadate at limiting concentrations of dissolved inorganic carbon indicated that more ATP was hydrolysed in algae grown at 70 muL . L-1 than in those grown at 400 muL . L-1 CO2. We conclude that the maximal capacity to accumulate dissolved inorganic carbon is inversely related to the CO2 concentration in the medium. Activation and - or synthesis of vanadate-sensitive ATPase may be the major explanation for the higher capacity for HCO3--dependent O2 evolution in cells grown under limited CO2 concentrations.