Search

Diatoms and other phytoplankton groups are exposed to abrupt changes in pCO2, in waters in upwelling areas, near CO2 seeps, or during their blooms; or to more gradual pCO2 rise through anthropogenic CO2 emissions. Gradual CO2 rises have, however, rarely been included in ocean acidification (OA) studies. We therefore compared how small (Thalassiosira pseudonana) and larger (Thalassiosira weissflogii) diatom cell isolates respond to gradual pCO2 rises from 180 to 1000 μatm in steps of ~40 μatm with 5–10 generations at each step, and whether their responses to gradual pCO2 rise differ when compared to an abrupt pCO2 rise imposed from ambient 400 directly to 1000 μatm. Cell volume increased in T. pseudonana but decreased in T. weissflogii with an increase from low to moderate CO2 levels, and then remained steady under yet higher CO2 levels. Growth rates were stimulated, but Chl a, particulate organic carbon (POC) and cellular biogenic silica (BSi) decreased from low to moderate CO2 levels, and then remained steady with further CO2 rise in both species. Decreased saturation light intensity (Ik) and light use efficiency (α) with CO2 rise in T. pseudonana indicate that the smaller diatom becomes more susceptible to photoinhibition. Decreased BSi/POC (Si/C) in T. weissflogii indicates the biogeochemical cycles of both silicon and carbon may be more affected by elevated pCO2 in the larger diatom. The different CO2 modulation methods resulted in different responses of some key physiological parameters. Increasing pCO2 from 180 to 400 μatm decreased cellular POC and BSi contents, implying that ocean acidification to date has already altered diatom contributions to carbon and silicon biogeochemical processes.

Light drives phytoplankton photosynthesis, so phytoplankton in their living habitats must exploit variable light levels and exposure durations, depending upon seasons, latitudes, depths and mixing events. Comparative growth, physiology and biochemical compositions were explored for the small Alexnadrium minutum (˜40 μm3 biovolume) and large Alexandrium catenella (˜9300 μm3 biovolume), globally wide spread coastal toxic red tide dinoflagellates, responding to a matrix of photoperiods (Light:Dark, 8:16, 16:8 and 24:0) and growth light irradiances. Smaller A. minutum grew faster under shorter photoperiods across growth light levels, while larger A. catenella grew fastest under longer photoperiods at the lowest applied light level. Photosystem II function responded largely to the instantaneous growth light level across photoperiod lengths, while the cell biovolume-based respiration, antioxidant capacity as well as cell composition responded more to photoperiod duration than to light level. These complex photophysiological responses resolved into linear correlations between growth rate versus cellular antioxidant activity and versus dark respiration, indicating that respiration energizes cellular antioxidant systems to benefit the growth of the cells. These results show the growth responses of Alexandrium species to light levels across photoperiods vary with species, and possibly with cell size. Together with previous results this puts a note of caution on meta-analytical extrapolations of physiological responses to light intensity derived from studies applying different photoperiods to different taxa, because different taxa show differential, even opposite growth responses to photoperiods and light intensities.

Summary • The lichen Lobaria pulmonaria survives large seasonal environmental changes through physiological acclimation to ambient conditions. • We quantitated algal cell population, cell division and key macromolecular levels associated with photosynthesis and nitrogen metabolism in L. pulmonaria sampled from four seasons with contrasting environmental conditions in a deciduous forest. • The algal symbiont population did not vary seasonally and cell division was restricted to the newest thallus margins. Nevertheless the symbiont concentrations of chlorophyll, PsbS, PsbA, and RbcL changed significantly through the seasons in the nondividing algal cells from older thallus regions. • L. pulmonaria reversibly allocated resources toward photochemical electron generation and carbohydrate production through the spring, summer and fall, and towards photoprotective dissipation in the cold, high-light winter. Our study shows that large seasonal molecular acclimation in L. pulmonaria occurs within a nearly stable, nondividing algal cell population that maintains photosynthetic capacity through many years of changing environmental cues.

Unicellular nitrogen fixing cyanobacteria (UCYN) are abundant members of phytoplankton communities in a wide range of marine environments, including those with rapidly changing nitrogen (N) concentrations. We hypothesized that differences in N availability (N2 vs. combined N) would cause UCYN to shift strategies of intracellular N and C allocation. We used transmission electron microscopy and nanoscale secondary ion mass spectrometry imaging to track assimilation and intracellular allocation of 13C-labeled CO2 and 15N-labeled N2 or NO3 at different periods across a diel cycle in Cyanothece sp. ATCC 51142. We present new ideas on interpreting these imaging data, including the influences of pre-incubation cellular C and N contents and turnover rates of inclusion bodies. Within cultures growing diazotrophically, distinct subpopulations were detected that fixed N2 at night or in the morning. Additional significant within-population heterogeneity was likely caused by differences in the relative amounts of N assimilated into cyanophycin from sources external and internal to the cells. Whether growing on N2 or NO3, cells prioritized cyanophycin synthesis when N assimilation rates were highest. N assimilation in cells growing on NO3 switched from cyanophycin synthesis to protein synthesis, suggesting that once a cyanophycin quota is met, it is bypassed in favor of protein synthesis. Growth on NO3 also revealed that at night, there is a very low level of CO2 assimilation into polysaccharides simultaneous with their catabolism for protein synthesis. This study revealed multiple, detailed mechanisms underlying C and N management in Cyanothece that facilitate its success in dynamic aquatic environments.

International audience; Over the seasons, Arctic diatom species occupy shifting habitats defined by contrasting light climates, constrained by snow and ice cover dynamics interacting with extreme photoperiod and solar angle variations. How Arctic diatom photoadaptation strategies differ across their heterogeneous light niches remains a poorly documented but crucial missing link to anticipate Arctic Ocean responses to shrinking sea-ice and increasing light. To address this question, we selected five Arctic diatom species with diverse life traits, representative of distinct light niches across the seasonal light environment continuum: from snow-covered dimly lit bottom ice to summer stratified waters. We studied their photoacclimation plasticity to two growth light levels and the subsequent responses of their nonphotochemical quenching (NPQ) and xanthophyll cycle to both dark incubations and light shifts. We deciphered NPQ and xanthophyll cycle tuning in darkness and their light-dependent induction kinet-ics, which aligned with species' light niche occupancy. In ice-related species, NPQ was sustained in darkness and its induction was more reactive to moderate light shifts. Open-water species triggered strong NPQ induction in darkness and reached higher maximal NPQ under high light. Marginal ice zone species showed strong adaptation to light fluctuations with a dark response fine-tuned depending upon light history. We argue these traits are anchored in diverging photoadaption strategies fostering Arctic diatom success in their respective light niches.

Significance Processes that darken the surface of the Greenland Ice Sheet (GrIS) enhance energy absorption and accelerate melt, with consequences for global sea-level rise. Here, we demonstrate how summer blooms of “glacier algae” darken the ice surface, significantly impacting the physical integrity of the environment. We identify and quantify the energy regulation mechanisms employed by glacier algae to balance their requirements for photosynthesis and growth with the extreme light and temperature regime of the GrIS, demonstrating how these mechanisms are optimized to darken and melt the ice surface. Our findings are critical for the incorporation of biological feedbacks into predictive models of GrIS surface runoff and provide unique insight into how photoautotrophic life excels within icy environments., Blooms of Zygnematophycean “glacier algae” lower the bare ice albedo of the Greenland Ice Sheet (GrIS), amplifying summer energy absorption at the ice surface and enhancing meltwater runoff from the largest cryospheric contributor to contemporary sea-level rise. Here, we provide a step change in current understanding of algal-driven ice sheet darkening through quantification of the photophysiological mechanisms that allow glacier algae to thrive on and darken the bare ice surface. Significant secondary phenolic pigmentation (11 times the cellular content of chlorophyll a) enables glacier algae to tolerate extreme irradiance (up to ∼4,000 µmol photons⋅m−2⋅s−1) while simultaneously repurposing captured ultraviolet and short-wave radiation for melt generation. Total cellular energy absorption is increased 50-fold by phenolic pigmentation, while glacier algal chloroplasts positioned beneath shading pigments remain low-light–adapted (Ek ∼46 µmol photons⋅m−2⋅s−1) and dependent upon typical nonphotochemical quenching mechanisms for photoregulation. On the GrIS, glacier algae direct only ∼1 to 2.4% of incident energy to photochemistry versus 48 to 65% to ice surface melting, contributing an additional ∼1.86 cm water equivalent surface melt per day in patches of high algal abundance (∼104 cells⋅mL−1). At the regional scale, surface darkening is driven by the direct and indirect impacts of glacier algae on ice albedo, with a significant negative relationship between broadband albedo (Moderate Resolution Imaging Spectroradiometer [MODIS]) and glacier algal biomass (R2 = 0.75, n = 149), indicating that up to 75% of the variability in albedo across the southwestern GrIS may be attributable to the presence of glacier algae.

Phytoplankton in the upper oceans are exposed to changing light levels due to mixing, diurnal solar cycles and weather conditions. Consequently, effects of ocean acidification are superimposed upon responses to variable light levels. We therefore grew a model diatom Thalassiosira pseudonana under either constant or variable light but at the same daily photon dose, with current low (400 μatm, LC) and future high CO2 (1000 μatm, HC) treatments. Variable light, compared with the constant light regime, decreased the growth rate, Chl a, Chl c, and carotenoid contents under both LC and HC conditions. Cells grown under variable light appeared more tolerant of high light as indicated by higher maximum relative electron transport rate and saturation light. Light variation interacted with high CO2/lowered pH to decrease the carbon fixation rate, but increased particulate organic carbon (POC) and particularly nitrogen (PON) per cell, which drove a decrease in C/N ratio, reflecting changes in the efficiency of energy transfer from photo-chemistry to net biomass production. Our results imply that elevated pCO2 under varying light conditions can lead to less primary productivity but more PON per biomass of the diatom, which might improve the food quality of diatoms and thereby influence biogeochemical nitrogen cycles.

Iron (Fe) is an essential cofactor for many metabolic enzymes of photoautotrophs. Although Fe limits phytoplankton productivity in broad areas of the ocean, phytoplankton have adapted their metabolism and growth to survive in these conditions. Using the euryhaline cyanobacterium Synechococcus sp. PCC7002, we investigated the physiological responses to long-term acclimation to four levels of Fe availability representative of the contemporary ocean (36.7, 3.83, 0.47 and 0.047 pM Fe’). With increasing severity of Fe limitation, Synechococcus sp. cells gradually decreased their volume and growth while increasing their energy allocation into organic carbon and nitrogen cellular pools. Furthermore, the total cellular content of pigments decreased. Additionally, with increasing severity of Fe limitation, intertwined responses of PSII functional cross-section (sPSII), re-oxidation time of the plastoquinone primary acceptor QA (t) and non-photochemical quenching revealed a shift in the photophysiological response between mild to strong Fe limitation compared with severe limitation. Under mild and strong Fe limitation, there was a decrease in linear electron transport accompanied by progressive loss of state transitions. Under severe Fe limitation, state transitions seemed to be largely supplanted by alternative electron pathways. In addition, mechanisms to dissipate energy excess and minimize oxidative stress associated with high irradiances increased with increasing severity of Fe limitation. Overall, our results establish the sequence of physiological strategies adopted by the cells under increasing severity of chronic Fe limitation, within a range of Fe concentrations relevant to modern ocean biogeochemistry.

Diatoms, as important contributors to aquatic primary production, are critical to the global carbon cycle. They tend to dominate phytoplankton communities experiencing rapid changes of underwater light. However, little is known regarding how climate change impacts diatoms’ capacity in coping with variable light environments. Here we grew a globally abundant diatom T. weissflogii, under two levels of temperature (18, 24 °C) and pCO2 (400, 1000 μatm), and then treated it with a light challenge to understand the combined effects of ocean warming and acidification on its exploitation of variable light environments. The higher temperature increased the photoinactivation rate at 400 μatm pCO2 and the higher pCO2 alleviated the negative effect of the higher temperature on PSII photoinactivation. Temperature did not affect the PsbA removal rate, but higher pCO2 stimulated PsbA removal. Photoinactivation outran repair, leading to decreased maximum photochemical yield in PSII. The higher pCO2 induced high sustained phase of nonphotochemical quenching when cells were less photoinhibited. The high light exposure induced the activity of both superoxide dismutase (SOD) and catalase (CAT) and the higher temperature stimulated them further, with insignificant effect of pCO2. Our findings suggest that ocean warming, ocean acidification and high light exposure would interact on PSII function and protection, and combination of these three environmental factors would lead to a reduced PSII activity in T. weissflogii. This study provides helpful insight into how climate change variables combined with local stressor impact diatoms’ photosynthetic physiology.

Fast Repetition and Relaxation chlorophyll fluorescence induction is used to estimate the effective absorption cross section of PSII (σPSII), to analyze phytoplankton acclimation and electron transport. The fitting coefficient ρ measures excitation transfer from closed PSII to remaining open PSII upon illumination, which could theoretically generate a progressive increase in σPSII for the remaining open PSII. To investigate how ρ responds to illumination we grew marine phytoplankters with diverse antenna structures (Prochlorococcus, Synechococcus, Ostreococcus and Thalassiosira pseudonana) under limiting or saturating growth light. Initial ρ varied with growth light in Synechococcus and Thalassiosira. With increasing actinic illumination PSII closed progressively and ρ decreased for all four taxa, in a pattern explicable as an exponential decay of ρ with increasing distance between remaining open PSII reaction centers. This light-dependent down-regulation of ρ allows the four phytoplankters to limit the effect of increasing light upon σPSII. The four structurally distinct taxa showed, however, distinct rates of response of ρ to PSII closure, likely reflecting differences in the spacing or orientation among their PSII centers. Following saturating illumination recovery of ρ in darkness coincided directly with PSII re-opening in Prochlorococcus. Even after PSII had re-opened in Synechococcus a transition to State II slowed dark recovery of ρ. In Ostreococcus sustained NPQ slowed dark recovery of ρ. In Thalassiosira dark recovery of ρ was slowed, possibly by a light-induced change in PSII spacing. These patterns of ρ versus PSII closure are thus a convenient probe of comparative PSII spacings.

Ocean acidification changes seawater chemistry, with increased CO2 and decreased pH regarded as the most important factors that impact marine organisms. This study employed an unconventional methodology to distinguish the independent effects of pH versus CO2. Changes in CO2 dominated the photochemical responses of the coastal diatom Phaeodactylum tricornutum to short-term ocean acidification. Increased CO2 lowered non-photochemical quenching of excitation and stimulated the electron transport rates of photosynthesis, with the largest effects on both parameters when CO2 and pH were altered simultaneously. Changes in pH alone did not show significant effects upon non-photochemical quenching (NPQ) nor upon electron transport rates, but can synergistically amplify CO2 effects under low light. Maximal induction of NPQ after illumination showed only a limited response to increasing CO2 under stable pH, across a range of increasing light levels, but maximal induced NPQ declined rapidly with increasing CO2 under variable pH, when measured under exposure to sub-saturating light, but not under saturating light. These findings show that aqueous CO2 and pH affect different physiological processes independently or interactively, which should be taken into account in future research for better understanding of responses to ocean acidification at the mechanistic level.

International audience

Picocyanobacteria are the numerically dominant photoautotrophs of the oligotrophic regions of Earth’s oceans. These organisms are characterized by their small size and highly reduced genomes. Strains partition to different light intensity and nutrient level niches, with differing photosynthetic apparatus stoichiometry, light harvesting machinery and susceptibility to photoinactivation. In this study, we grew three strains of picocyanobacteria: the low light, high nutrient strain Prochlorococcus marinus MIT 9313; the high light, low nutrient Prochlorococcus marinus MED 4; and the high light, high nutrient marine Synechococcus strain WH 8102; under low and high growth light levels. We then performed matched photophysiology, protein and transcript analyses. The strains differ significantly in their rates of Photosystem II repair under high light and in their capacity to remove the PsbA protein as the first step in the Photosystem II repair process. Notably, all strains remove the PsbD subunit at the same rate that they remove PsbA. When grown under low light, MIT 9313 loses active Photosystem II quickly when shifted to high light, but has no measurable capacity to remove PsbA. MED 4 and WH 8102 show less rapid loss of Photosystem II and considerable capacity to remove PsbA. MIT 9313 has less of the FtsH protease thought to be responsible for the removal of PsbA in other cyanobacteria. Furthermore, by transcript analysis the predominant FtsH isoform expressed in MIT 9313 is homologous to the FtsH 4 isoform characterized in the model strain Synechocystis PCC 6803, rather than the FtsH 2 and 3 isoforms thought to be responsible for PsbA degradation. MED 4 on the other hand shows high light inducible expression of the isoforms homologous to FtsH 2 and 3, consistent with its faster rate of PsbA removal. MIT 9313 has adapted to its low light environment by diverting resources away from Photosystem II content and repair.

Diatom responses to ocean acidification have been documented with variable and controversial results. We grew the coastal diatom Thalassiosira weissflogii under 410 (LC, pH 8.13) vs 1000 μatm (HC, pH 7.83) pCO2 and at different levels of light (80, 140, 220 μmol photons m−2 s−1), and found that light level alters physiological responses to OA. CO2 concentrating mechanisms (CCMs) were down-regulated in the HC-grown cells across all the light levels, as reflected by lowered activity of the periplasmic carbonic anhydrase and decreased photosynthetic affinity for CO2 or dissolved inorganic carbon. The specific growth rate was, however, enhanced significantly by 9.2% only at the limiting low light level. These results indicate that rather than CO2 “fertilization”, the energy saved from down-regulation of CCMs promoted the growth rate of the diatom when light availability is low, in parallel with enhanced respiration under OA to cope with the acidic stress by providing extra energy.

Marine phytoplankton, and in particular diatoms, are responsible for almost half of all primary production on Earth. Diatom species thrive from polar to tropical waters and across light environments that are highly complex to relatively benign, and so have evolved highly divergent strategies for regulating light capture and utilization. It is increasingly well established that diatoms have achieved such successful ecosystem dominance by regulating excitation energy available for generating photosynthetic energy via highly flexible light harvesting strategies. However, how different light harvesting strategies and downstream pathways for oxygen production and consumption interact to balance excitation pressure remains unknown. We therefore examined the responses of three diatom taxa adapted to inherently different light climates (estuarine Thalassioisira weissflogii, coastal Thalassiosira pseudonana and oceanic Thalassiosira oceanica) during transient shifts from a moderate to high growth irradiance (85 to 1200 μmol photons m-2 s-1). Transient high light exposure caused T. weissflogii to rapidly downregulate PSII with substantial nonphotochemical quenching, protecting PSII from inactivation or damage, and obviating the need for induction of O2 consuming (light-dependent respiration, LDR) pathways. In contrast, T. oceanica retained high excitation pressure on PSII, but with little change in RCII photochemical turnover, thereby requiring moderate repair activity and greater reliance on LDR. T. pseudonana exhibited an intermediate response compared to the other two diatom species, exhibiting some downregulation and inactivation of PSII, but high repair of PSII and induction of reversible PSII nonphotochemical quenching, with some LDR. Together, these data demonstrate a range of strategies for balancing light harvesting and utilization across diatom species, which reflect their adaptation to sustain photosynthesis under environments with inherently different light regimes.

Light drives phytoplankton productivity, so phytoplankton must exploit variable intensities and durations of light exposure, depending upon season, latitude, and depth. We analyzed the growth, photophysiology and composition of small, Thalassiosira pseudonana, and large, Thalassiosira punctigera, centric diatoms from temperate, coastal marine habitats, responding to a matrix of photoperiods and growth light intensities. T. pseudonana showed fastest growth rates under long photoperiods and low to moderate light intensities, while the larger T. punctigera showed fastest growth rates under short photoperiods and higher light intensities. Photosystem II function and content responded primarily to instantaneous growth light intensities during the photoperiod, while diel carbon fixation and RUBISCO content responded more to photoperiod duration than to instantaneous light intensity. Changing photoperiods caused species-specific changes in the responses of photochemical yield (e- /photon) to growth light intensity. These photophysiological variables showed complex responses to photoperiod and to growth light intensity. Growth rate also showed complex responses to photoperiod and growth light intensity. But these complex responses resolved into a close relation between growth rate and the cumulative daily generation of reductant, across the matrix of photoperiods and light intensities.

The title of Beth Shapiro’s ‘How to Clone a Mammoth’ contains an implicature: it suggests that it is indeed possible to clone a mammoth, to bring extinct species back from the dead. But in fact Shapiro both denies this is possible, and denies there would be good reason to do it even if it were possible. The de-extinct ‘mammoths’ she speaks of are merely ecological proxies for mammoths—elephants re-engineered for cold-tolerance by the addition to their genomes of a few mammoth genes. Shapiro’s denial that genuine species de-extinction is possible is based on her assumption that resurrected organisms would need to be perfectly indistinguishable from the creatures that died out. In this article I use the example of an extinct New Zealand wattlebird, the huia, to argue—contra Shapiro—that there are compelling reasons to resurrect certain species if it can be done. I then argue—again, contra Shapiro—that synthetically created organisms needn’t be perfectly indistinguishable from their genetic forebears in order for species de-extinction to be successful.

International audience; The effective absorption cross section for photochemistry of Photosystem II in the light (σ PSII ′) comprises the probability of light capture by Photosystem II and the quantum yield for subsequent photochemistry. σ PSII ′ is used to model photosynthesis and aquatic productivity, and phytoplankters regulate σ PSII ′ to mitigate over-or under-excitation of Photosystem II. We used diverse phytoplankton taxa to compare short and long term changes in σ PSII ′ with the induction of the yield of non-photochemical quenching (YNPQ) of chlorophyll fluorescence, a measure of regulated excitation dissipation. In two picocyanobacteria σ PSII ′ showed no decline upon induction of moderate YNPQ, above light levels sufficient for saturation of electron transport. In the eukaryotic chl a/b Ostreococcus and the chl a/c diatom Thalassiosira, induction of non-photochemical quenching was stronger after growth under saturating light, an acclimation attributable to increased xanthophyll cycle pigment content. Across short and longer-term light histories to induce or relax regulatory processes Ostreococcus and Thalassiosira showed proportional variations between the level of YNPQ and the down regulation of σ PSII ′. The proportional down regulation of σ PSII ′ was, however, significantly smaller than the amplitude of YNPQ induction. For the eukaryotes we can predict changes in σ PSII ′ , useful for modeling electron transport, productivity and acclimation, from measures of YNPQ, which are accessible from fluorescence yield measures that do not include σ PSII ′. This useful relation, however, does not extend to the tested prokaryotes, possibly as a result of differential violations of the rate constant assumptions that underlie the calculated YNPQ parameter.

Nitrogen stress is an important control on the growth of phytoplankton and varying responses to this common condition among taxa may affect their relative success within phytoplankton communities. We analyzed photosynthetic responses to nitrogen (N) stress in two classes of phytoplankton that often dominate their respective size ranges, diatoms and prasinophytes, selecting species of distinct niches within each class. Changes in photosynthetic structures appeared similar within each class during N stress, but photophysiological and growth responses were more species- or niche-specific. In the coastal diatom Thalassiosira pseudonana and the oceanic diatom T. weissflogii, N starvation induced large declines in photosynthetic pigments and Photosystem II (PSII) quantity and activity as well as increases in the effective absorption cross-section of PSII photochemistry (σʹPSII). These diatoms also increased photoprotection through energy-dependent non-photochemical quenching (NPQ) during N starvation. Resupply of N in diatoms caused rapid recovery of growth and relaxation of NPQ, while recovery of PSII photochemistry was slower. In contrast, the prasinophytes Micromonas sp., an Arctic Ocean species, and Ostreococcus tauri, a temperate coastal eutrophile, showed little change in photosynthetic pigments and structures and a decline or no change, respectively, in σʹPSII with N starvation. Growth and PSII function recovered quickly in Micromonas sp. after resupply of N while O. tauri failed to recover N-replete levels of electron transfer from PSII and growth, possibly due to their distinct photoprotective strategies. O. tauri induced energy-dependent NPQ for photoprotection that may suit its variable and nutrient-rich habitat. Micromonas sp. relies upon both energy-dependent NPQ and a sustained, energy-independent NPQ mechanism. A strategy in Micromonas sp. that permits photoprotection with little change in photosynthetic structures is consistent with its Arctic niche, where low temperatures and thus low biosynthetic rates create higher opportunity costs to rebuild photosynthetic structures.

Micromonas strains of small prasinophyte green algae are found throughout the world's oceans, exploiting widely different niches. We grew arctic and temperate strains of Micromonas and compared their susceptibilities to photoinactivation of Photosystem II, their counteracting Photosystem II repair capacities, their Photosystem II content, and their induction and relaxation of non-photochemical quenching. In the arctic strain Micromonas NCMA 2099, the cellular content of active Photosystem II represents only about 50 % of total Photosystem II protein, as a slow rate constant for clearance of PsbA protein limits instantaneous repair. In contrast, the temperate strain NCMA 1646 shows a faster clearance of PsbA protein which allows it to maintain active Photosystem II content equivalent to total Photosystem II protein. Under growth at 2 °C, the arctic Micromonas maintains a constitutive induction of xanthophyll deepoxidation, shown by second-derivative whole-cell spectra, which supports strong induction of non-photochemical quenching under low to moderate light, even if xanthophyll cycling is blocked. This non-photochemical quenching, however, relaxes during subsequent darkness with kinetics nearly comparable to the temperate Micromonas NCMA 1646, thereby limiting the opportunity cost of sustained downregulation of PSII function after a decrease in light.

Marine Synechococcus and Prochlorococcus are picocyanobacteria predominating in subtropical, oligotrophic marine environments, a niche predicted to expand with climate change. When grown under common low light conditions Synechococcus WH 8102 and Prochlorococcus MED 4 show similar Cytochrome b6f and Photosystem I contents normalized to Photosystem II content, while Prochlorococcus MIT 9313 has twice the Cytochrome b6f content and four times the Photosystem I content of the other strains. Interestingly, the Prochlorococcus strains contain only one third to one half of the RUBISCO catalytic subunits compared to the marine Synechococcus strain. The maximum Photosystem II electron transport rates were similar for the two Prochlorococcus strains but higher for the marine Synechococcus strain. Photosystem II electron transport capacity is highly correlated to the molar ratio of RUBISCO active sites to Photosystem II but not to the ratio of cytochrome b6f to Photosystem II, nor to the ratio of Photosystem I: Photosystem II. Thus, the catalytic capacity for the rate-limiting step of carbon fixation, the ultimate electron sink, appears to limit electron transport rates. The high abundance of Cytochrome b6f and Photosystem I in MIT 9313, combined with the slower flow of electrons away from Photosystem II and the relatively low level of RUBISCO, are consistent with cyclic electron flow around Photosystem I in this strain.

The diazotrophic cyanobacterium Trichodesmium is a major contributor to marine nitrogen fixation. We analyzed how light acclimation influences the photophysiological performance of Trichodesmium IMS101 during exponential growth in semi-continuous nitrogen fixing cultures under light levels of 70, 150, 250, and 400 μmol photons m−2 s−1, across diel cycles. There were close correlations between growth rate, trichome length, particulate organic carbon and nitrogen assimilation, and cellular absorbance, which all peaked at 150 μmol photons m−2 s−1. Growth rate was light saturated by about 100 μmol photons m−2 s−1 and was photoinhibited above 150 μmol photons m−2 s−1. In contrast, the light level (I k) to saturate PSII electron transport (e − PSII−1 s−1) was much higher, in the range of 450–550 μmol photons m−2 s−1, and increased with growth light. Growth rate correlates with the absorption cross section as well as with absorbed photons per cell, but not to electron transport per PSII; this disparity suggests that numbers of PSII in a cell, along with the energy allocation between two photosystems and the state transition mechanism underlie the changes in growth rates. The rate of state transitions after a transfer to darkness increased with growth light, indicating faster respiratory input into the intersystem electron transport chain.

Diatoms are marine primary producers that sink in part due to the density of their silica frustules. Sinking of these phytoplankters is crucial for both the biological pump that sequesters carbon to the deep ocean and for the life strategy of the organism. Sinking rates have been previously measured through settling columns, or with fluorimeters or video microscopy arranged perpendicularly to the direction of sinking. These side-view techniques require large volumes of culture, specialized equipment and are difficult to scale up to multiple simultaneous measures for screening. We established a method for parallel, large scale analysis of multiple phytoplankton sinking rates through top-view monitoring of chlorophyll a fluorescence in microtitre well plates. We verified the method through experimental analysis of known factors that influence sinking rates, including exponential versus stationary growth phase in species of different cell sizes; Thalassiosira pseudonana CCMP1335, chain-forming Skeletonema marinoi RO5A and Coscinodiscus radiatus CCMP312. We fit decay curves to an algebraic transform of the decrease in fluorescence signal as cells sank away from the fluorometer detector, and then used minimal mechanistic assumptions to extract a sinking rate (m d-1) using an RStudio script, SinkWORX. We thereby detected significant differences in sinking rates as larger diatom cells sank faster than smaller cells, and cultures in stationary phase sank faster than those in exponential phase. Our sinking rate estimates accord well with literature values from previously established methods. This well plate-based method can operate as a high throughput integrative phenotypic screen for factors that influence sinking rates including macromolecular allocations, nutrient availability or uptake rates, chain-length or cell size, degree of silification and progression through growth stages. Alternately the approach can be used to phenomically screen libraries of mutants.

© 2016 The Authors Limnology and Oceanography: Methods published by Wiley Periodicals, Inc. Photosystem II (PSII) is a pigment-protein complex that photochemically extracts electrons from water, generating the reductant that supports biological productivity in all biomes. Estimating the content of active PSII reaction centers in a liquid sample is a key input for estimating aquatic photosynthesis rates, as well as for analyzing phytoplankton stress responses. Established procedures for PSII content quantification based on oxygen evolution are slow, imprecise and require dense cell suspensions, and are thus inapplicable to many laboratory or field studies. A new approach uses baseline chlorophyll fluorescence emission divided by the effective absorbance cross section for PSII photochemistry, with both variables derivable from single turnover fluorescence induction protocols. This approach has not been widely tested and is potentially subject to variation in samples suffering progressive photoinactivation or induction of non-photochemical quenching under variable light. We evaluated the validity of this approach for a marine picocyanobacteria, low and high light Prochlorococcus ecotypes, arctic and temperate prasinophyte green alga and two centric diatoms, generating 209 paired determinations from a range of growth and treatment conditions. We successfully calibrated the fluorescence derived estimator for PSII reaction center content, and demonstrate a modification that corrects for the short term influence of photoinactivation. The modified parameter shows little response to induction of non-photochemical quenching. In doing so we show the potential and limitations of an estimator of active PSII reaction center content that is sufficiently robust to support rapid, time-resolved autonomous measures of primary productivity from lakes and oceans.

This chapter examines three prime candidates for de-extinction—namely, the aurochs, the woolly mammoth, and the passenger pigeon. It will be about what these animals were like, why people want to resurrect them, and the methods by which their resurrections could be accomplished.

Diatoms are the most important group of primary producers in marine ecosystems. As oceanic pH declines and increased stratification leads to the upper mixing layer becoming shallower, diatoms are interactively affected by both lower pH and higher average exposures to solar ultraviolet radiation. The photochemical yields of a model diatom, Phaeodactylum tricornutum, were inhibited by ultraviolet radiation under both growth and excess light levels, while the functional absorbance cross sections of the remaining photosystem II increased. Cells grown under ocean acidification (OA) were less affected during UV exposure. The recovery of PSII under low photosynthetically active radiation was much faster than in the dark, indicating that photosynthetic processes were essential for the full recovery of photosystem II. This light dependent recovery required de novo synthesized protein. Cells grown under ocean acidification recovered faster, possibly attributable to higher CO2 availability for the Calvin cycle producing more resources for repair. The lower UV inhibition combined with higher recovery rate under ocean acidification could benefit species such as P. tricornutum, and change their competitiveness in the future ocean.

Ocean acidification is changing the nature of inorganic carbon availability in the global oceans. Diatoms account for , 40% of all marine primary productivity and are major contributors to the export of atmospheric carbon to the deep ocean. Larger diatoms are more likely to be stimulated by future increases in CO2 availability as a result of their low surface area to volume ratio and lower diffusive flux of CO2 relative to their carbon demand for growth. Here we quantify the effect of the partial pressure of carbon dioxide (PCO2), at levels of 190, 380, and 750 m LL 21, on the growth rate, photosystem II electron transport rate (ETR), and elemental composition for five diatom species ranging over five orders of magnitude in cell volume. Growth rates for all species were enhanced under 750 relative to 190 and 380 m LL 21, with little change in ETR or elemental stoichiometries, indicating an enhanced allocation of photochemical energy to growth under elevated PCO2 .P CO2 enhancement of growth rates was size dependent. Under 750 vs. 190 m LL 21 partial pressures, growth rate was enhanced by , 5% for the smaller diatom species to , 30% for the largest species examined. The size dependence of CO2-stimulated growth enhancement indicates that ocean acidification may selectively favor an increase in the growth rates of larger vs. smaller phytoplankton species in the sea, with potentially significant consequences for carbon biochemistry.

Changing light environments force photoautotroph cells, including coral symbionts, to acclimate to maintain photosynthesis. Photosystem II (PSII) is subjected to photoinactivation at a rate proportional to the incident light, and cells must adjust their rates of protein repair to counter this photoinactivation. We examined PSII function in the coral symbiont Symbiodinium to determine the effect of photoacclimation on their capacity for PSII repair. Colonies of the coral Stylophora pistillata were collected from moderate light environments on the Lizard Island reef (Queensland, Australia) and transported to a local field station, where they were assigned to lower or higher light regimes and allowed to acclimate for 2 weeks. Following this photoacclimation period, the low-light acclimated corals showed greater symbiont density, higher chlorophyll per symbiont cell, and higher photosystem II protein than high-light acclimated corals did. Subsequently, we treated the corals with lincomycin, an inhibitor of chloroplastic protein synthesis, and exposed them to a high-light treatment to separate the effect of de novo protein synthesis in PSII repair from intrinsic susceptibility to photoinactivation. Low-light acclimated corals showed a sharp initial drop in PSII function but inhibition of PSII repair provoked only a modest additional drop in PSII function, compared to uninhibited corals. In high-light acclimated corals inhibition of PSII repair provoked a larger drop in PSII function, compared to uninhibited high-light corals. The greater lincomycin effects in the corals pre-acclimated to high-light show that high-light leads to an increased reliance on the PSII repair cycle.

All oxygenic photoautotrophs suffer photoinactivation of their Photosystem II complexes, at a rate driven by the instantaneous light level. To maintain photosynthesis, PsbA subunits are proteolytically removed from photoinactivated Photosystem II complexes, primarily by a membrane-bound FtsH protease. Diatoms thrive in environments with fluctuating light, such as coastal regions, in part because they enjoy a low susceptibility to photoinactivation of Photosystem II. In a coastal strain of the diatom Thalassiosira pseudonana growing across a range of light levels, active Photosystem II represents only about 42 % of the total Photosystem II protein, with the remainder attributable to photoinactivated Photosystem II awaiting recycling. The rate constant for removal of PsbA protein increases with growth light, in parallel with an increasing content of the FtsH protease relative to the substrate PsbA. An offshore strain of Thalassiosira pseudonana, originating from a more stable light environment, had a lower content of FtsH and slower rate constants for removal of PsbA. We used this data to generate the first estimates for in vivo proteolytic degradation of photoinactivated PsbA per FtsH(6) protease, at similar to 3.9 x 10(-2) s(-1), which proved consistent across growth lights and across the onshore and offshore strains.

Marine diatoms are photosynthetic, and thrive in environments where light fluctuates. Like all oxygenic photosynthetic organisms diatoms face a light-dependent inactivation of the Photosystem II complexes that photooxidize water to generate biosynthetic reductant. To maintain photosynthesis this photoinactivation must be countered by slow and metabolically expensive protein turnover, which is light dependent in cyanobacteria and in plants. We tracked daily cycles of the content, synthesis and degradation of Photosystem II, in a small and in a large marine diatom, under low and high growth light levels. We show that, unlike plants, diatoms maintain extensive cycling of Photosystem II proteins even in the dark. Photosystem II protein cycling saturates at low light, and continued cycling in dark periods, using energy from respiration, allows the diatoms to catch up to excess photoinactivation accumulated over the preceding illuminated period. The large diatom suffers only limited photoinactivation of Photosystem II, but cycling of Photosystem II protein exceeds Photosystem II inactivation, so the large diatom recycles functional Photosystem II units before they are inactivated. Through the diel cycle the contents of active Photosystem II centers and Photosystem II proteins change predictably, but are not correlated, generating large changes in the fraction of total PSII that is active at a given time or growth condition. We propose that dark and steady cycling of Photosystem II proteins is driven by the tight integration of chloroplastic and mitochondrial metabolism in diatoms. This ability for baseline, continuous Photosystem II repair could contribute to the success of diatoms in mixed water environments that carry them from illumination to darkness and back.

The in hospite Symbiodinium symbiont of corals on shallow reefs relies on photoprotection and photorepair during periods of exposure to short-term high light and/or temperature stress. A coral’s susceptibility to bleaching is species specific and determined not only by Symbiodinium type, size and physiology, but also by coral host features. Here, photoprotective, photorepair, photochemical and non-photochemical efficiency parameters of Symbiodinium harboured in two morphologically different coral species were examined on Heron Island (23.4420°S, 151.9140°E) in July 2011. The two coral species were exposed to high light stress for 96 h, with or without inhibition of photosystem (PS) II repair by lincomycin. Symbiodinium harboured in Pocillopora damicornis showed an increase in xanthophyll de-epoxidation under high light exposure, whereas algal symbionts in Pavona decussata showed constant levels of xanthophyll de-epoxidation. High light-treated specimens of P. damicornis maintained steady PsbA protein (D1 protein) content throughout the experiment, but P. decussata showed a peak in PsbA protein content after 48 h of exposure. In hospite Symbiodinium in P. damicornis had greater content of PsbA protein fragments, suggesting higher accumulation of photodamaged products, compared to Symbiodinium in P. decussata, where both maintained steady PSII photochemical capacity over 96 h of exposure. Under inhibition of PSII repair, both species lost PsbA protein content and PSII photochemical capacity. Both species showed increased heat dissipation under inhibition of PSII repair, but differed in photoprotective strategies and photorepair activity. Our results suggest that, as well as any differences in the symbiont, characteristics of the coral host can alter important physiological responses in Symbiodinium.

International audience; Skeletonema costatum and Phaeodactylum tricornutum are model marine diatoms with differing strategies for non-photochemical dissipation of excess excitation energy within photosystem II (PSII). We showed that S. costatum, with connectivity across the pigment bed serving PSII, and limited capacity for induction of sustained non-photochemical quenching (NPQ), maintained a large ratio of [PSIITotal]/[PSIIActive] to buffer against fluctuations in light intensity. In contrast, P. tricornutum, with a larger capacity to induce sustained NPQ, could maintain a lower [PSIITotal]/[PSIIActive]. Induction of NPQ was correlated with an active PSII repair cycle in both species, and inhibition of chloroplastic protein synthesis with lincomycin leads to run away over-excitation of remaining PSIIActive, particularly in S. costatum. We discuss these distinctions in relation to the differing capacities, induction and relaxation rates for NPQ, and as strain adaptations to the differential light regimes of their originating habitats. The present work further confirms the important role for the light-dependent fast regulation of photochemistry by NPQ interacting with PSII repair cycle capacity in the ecophysiology of both pennate and centric diatoms.

Increasing anthropogenic carbon dioxide is causing changes to ocean chemistry, which will continue in a predictable manner. Dissolution of additional atmospheric carbon dioxide leads to increased concentrations of dissolved carbon dioxide and bicarbonate and decreased pH in ocean water. The concomitant effects on phytoplankton ecophysiology, leading potentially to changes in community structure, are now a focus of concern. Therefore, we grew the coccolithophore Emiliania huxleyi (Lohmann) W. W. Hay et H. Mohler and the diatom strains Thalassiosira pseudonana (Hust.) Hasle et Heimdal CCMP 1014 and T. pseudonana CCMP 1335 under low light in turbidostat photobioreactors bubbled with air containing 390 ppmv or 750 ppmv CO2 . Increased pCO2 led to increased growth rates in all three strains. In addition, protein levels of RUBISCO increased in the coastal strains of both species, showing a larger capacity for CO2 assimilation at 750 ppmv CO2 . With increased pCO2 , both T. pseudonana strains displayed an increased susceptibility to PSII photoinactivation and, to compensate, an augmented capacity for PSII repair. Consequently, the cost of maintaining PSII function for the diatoms increased at increased pCO2 . In E. huxleyi, PSII photoinactivation and the counter-acting repair, while both intrinsically larger than in T. pseudonana, did not change between the current and high-pCO2 treatments. The content of the photosynthetic electron transport intermediary cytochrome b6/f complex increased significantly in the diatoms under elevated pCO2 , suggesting changes in electron transport function.

This essay summarizes my book The Deliverance of God: An Apocalyptic Rereading of Justification in Paul in two main stages. First it describes the book’s distinctive account of a problem in modern Pauline scholarship in relation to several key debates and some of their most important representative figures—Wrede, Sanders, Stendahl, and Martyn. It then describes how the book offers in its first half (Parts I-III) an underlying unified and theological account of these issues—the unwitting release of Arianism within Paul’s interpretation in the specific form of a conditional conception of salvation in terms of a sequence of contracts. Following this the essay charts quickly the solution to this conundrum offered by the book’s second half (Parts IV and V): a non-contractual reading of all the texts in Paul that could generate a problematic contractual and conditional construal. This rereading concentrates on Romans 1-4 (1:16-5:1), and, within that passage, on the especially important 1:18-3:20, which is construed as a Socratic argument and thereby unconditionally in the broader setting of Romans.

This is a rejoinder to the criticisms of Grant Macaskill and Barry Matlock of The Deliverance of God. Because they largely misunderstand the complex argument of Deliverance it is redescribed and exemplified here in relation to Gal. 2.15-16, in the light of which the concerns of Macaskill and Matlock can be seen to be either misdirected or inadequate. Counter to their negative contentions it is argued that the essentially Arian problems within Paul’s description described by Deliverance remain real, widespread, and critical, but that no serious difficulties are visible as yet in the fundamentally Athanasian reading of Paul’s justification texts Deliverance offers. In alternative terms: if Paul is read consistently and rigorously ‘backward’, and never ‘forward’, a quagmire of interpretative difficulties can be negotiated, but not otherwise.

Diatoms host chlorophyll a/c chloroplasts distinct from green chloroplasts. Diatoms now dominate the eukaryotic oceanic phytoplankton, in part through their exploitation of environments with variable light. We grew marine diatoms across a range of temperatures and then analyzed their PSII function and subunit turnover during an increase in light to mimic an upward mixing event. The small diatom Thalassiosira pseudonana initially responds to increased photoinactivation under blue or white light with rapid acceleration of the photosystem II (PSII) repair cycle. Increased red light provoked only modest PSII photoinactivation but triggered a rapid clearance of a subpool of PsbA. Furthermore, PsbD and PsbB content was greater than PsbA content, indicating a large pool of partly assembled PSII repair cycle intermediates lacking PsbA. The initial replacement rates for PsbD (D2) were, surprisingly, comparable to or higher than those for PsbA (D1), and even the supposedly stable PsbB (CP47) dropped rapidly upon the light shift, showing a novel aspect of rapid protein subunit turnover in the PSII repair cycle in small diatoms. Under sustained high light, T. pseudonana induces sustained nonphotochemical quenching, which correlates with stabilization of PSII function and the PsbA pool. The larger diatom Coscinodiscus radiatus showed generally similar responses but had a smaller allocation of PSII complexes relative to total protein content, with nearly equal stiochiometries of PsbA and PsbD subunits. Fast turnover of multiple PSII subunits, pools of PSII repair cycle intermediates, and photoprotective induction of nonphotochemical quenching are important interacting factors, particularly for small diatoms, to withstand and exploit high, fluctuating light.

We studied the interactions of the CO2-concentrating mechanism and variable light in the filamentous cyanobacterium Leptolyngbya sp. CPCC 696 acclimated to low light (15 μmol m−2 s−1 PPFD) and low inorganic carbon (50 μM Ci). Mass spectrometric and polarographic analysis revealed that mediated CO2 uptake along with both active Na+-independent and Na+-dependent HCO3 − transport, likely through Na+/HCO3 − symport, were employed to concentrate Ci internally. Combined transport of CO2 and HCO3 − required about 30 kJ mol−1 of energy from photosynthetic electron transport to support an intracellular Ci accumulation 550-fold greater than the external Ci. Initially, Leptolyngbya rapidly induced oxygen evolution and Ci transport to reach 40–50% of maximum values by 50 μmol m−2 s−1 PPFD. Thereafter, photosynthesis and Ci transport increased gradually to saturation around 1,800 μmol m−2 s−1 PPFD. Leptolyngbya showed a low intrinsic susceptibility to photoinhibition of oxygen evolution up to PPFD of 3,000 μmol m−2 s−1. Intracellular Ci accumulation showed a lag under low light but then peaked at about 500 μmol photons m−2 s−1 and remained high thereafter. Ci influx was accompanied by a simultaneous, light-dependent, outward flux of CO2 and by internal CO2/HCO3 − cycling. The high-affinity and high-capacity CCM of Leptolyngbya responded dynamically to fluctuating PPFD and used excitation energy in excess of the needs of CO2 fixation by increasing Ci transport, accumulation and Ci cycling. This capacity may allow Leptolyngbya to tolerate periodic exposure to excess high light by consuming electron equivalents and keeping PSII open.

We dissect the primary photo-inactivation and the counteracting metabolic repair rates in fragments of the scleractinian coral, Pocillopora damicornis, subjected to a combined stress of a shift to elevated temperature (from 26°C to 32°C) and increased light (from 200 μmol photons m-2 s-1 to 400 μmol photons m-2 s-1) to induce bleaching. During the bleaching treatment the dinoflagellate symbionts showed a 5.5-fold acceleration in their photosystem II (PSII) repair rate constant, demonstrating that they maintain strong metabolic capacity to clear and replace photo-damaged D1 protein at the elevated temperature and light conditions. Nevertheless, the symbionts concurrently suffered a seven-fold increase in the rate constant for PSII photo-inactivation. This rapid photo-inactivation exceeded the PSII repair capacity, therefore tipping the symbionts, and by implication the symbiosis, into net photo-inhibition. Increased photo-inactivation in hospite, rather than an inhibition of PSII repair, is the principle trigger for net photo-inhibition under bleaching conditions. © 2011, by the American Society of Limnology and Oceanography, Inc.

We quantified and compared the photophysiological characteristics of Emiliania huxleyi with two marine diatoms Thalassiosira pseudonana and Coscinodiscus sp. under nitrogen- and phosphorus-replete and -depleted conditions. Under nitrogen-depleted conditions, E. huxleyi maintained significant photosystem II (PSII) function over at least 38 d, whereas for both diatoms, PSII function declined to marginal levels within 8–10 d of nitrogen depletion. In contrast, under phosphorus-depleted conditions, PSII function in E. huxleyi declined sharply within 7 d and in T. pseudonana within 3 d. Under nutrient-replete conditions, E. huxleyi had one of the highest PSII repair rate constants among phytoplankton examined to date. Under nitrogen depletion, E. huxleyi exhibited a decrease in susceptibility to photoinactivation of PSII, while the diatoms showed no significant change in susceptibility. The superior ability of E. huxleyi relative to the diatoms to limit PSII photoinactivation as well as to maintain PSII repair for many days of nitrogen depletion could help to explain why E. huxleyi is able to form blooms under the low-nitrate, high-light conditions that frequently occur in stratified surface waters.

We examined the influence of forecasted changes in global temperatures and pCO2 on N2 fixation and assimilation in the ecologically important cyanobacterium Trichodesmium spp. Changes of mRNA transcripts (nifH, glnA, hetR, psbA, psaB), protein (nitrogenase, glutamine synthetase) pools and enzymatic activity (nitrogenase) were measured under varying pCO2 and temperatures. High pCO2 shifted transcript patterns of all genes, resulting in a more synchronized diel expression. Under the same conditions, we did not observe any significant changes in the protein pools or in total cellular allocations of carbon and nitrogen (i.e. C : N ratio remained stable). Independently of temperature, high pCO2 (900 µatm) elevated N2 fixation rates. Levels of the key enzymes, nitrogenase and glutamine synthetase that mediate nitrogen assimilation did not increase, implying that the high pCO2 allowed higher reaction turnover rates through these key enzymes. Moreover, increased temperatures and high pCO2 resulted in higher C : P ratios. The plasticity in phosphorous stoichiometry combined with higher enzymatic efficiencies lead to higher growth rates. In cyanobacteria photosynthesis, carbon uptake, respiration, N2 fixation and nitrogen assimilation share cellular components. We propose that shifted cellular resource and energy allocation among those components will enable Trichodesmium grown at elevated temperatures and pCO2 to extend its niche in the future ocean, through both tolerance of a broader temperature range and higher P plasticity.

International audience; We investigated the photophysiological responses of three ecotypes of the picophytoplankter Ostreococcus and a larger prasinophyte Pyramimonas obovata to a sudden increase in light irradiance. The deepwater Ostreococcus sp. RCC809 showed very high susceptibility to primary photoinactivation, likely a consequence of high oxidative stress, which may relate to the recently noted plastid terminal oxidase activity in this strain. The three Ostreococcus ecotypes were all capable of deploying modulation of the photosystem II repair cycle in order to cope with the light increase, but the effective clearance of photoinactivated D1 protein appeared to be slower in the deepwater Ostreococcus sp. RCC809, suggesting that this step is rate limiting in the photosystem II repair cycle in this strain. Moreover, the deepwater Ostreococcus accumulated lutein and showed substantial use of the xanthophyll cycle under light stress, demonstrating its high sensitivity to light fluctuations. The sustained component of the nonphotochemical quenching of fluorescence correlated well with the xanthophyll deepoxidation activity. Comparisons with the larger prasinophyte P. obovata suggest that the photophysiology of Ostreococcus ecotypes requires high photosystem II repair rates to counter a high susceptibility to photoinactivation, consistent with low pigment package effects in their minute-sized cells.

International audience; In cyanobacteria, the D1 protein of photosystem II (PSII) is encoded by the psbA multigene family. In most freshwater strains, a D1:1 isoform of this protein is exchanged for a D1: 2 isoform in response to various stresses, thereby altering PSII photochemistry. To investigate PSII responses to stress in marine Synechococcus, we acclimated cultures of the WH7803 strain to different growth irradiances and then exposed them to high light (HL) or ultraviolet (UV) radiation. Measurement of PSII quantum yield and quantitation of the D1 protein pool showed that HL-acclimated cells were more resistant to UV light than were low light-(LL) or medium light-(ML) acclimated cells. Both UV and HL induced the expression of psbA genes encoding D1: 2 and the repression of the psbA gene encoding D1: 1. Although three psbA genes encode identical D1: 2 isoforms in Synechococcus sp. WH7803, only one was strongly stress responsive in our treatment conditions. Examination of 11 marine Synechococcus genomic sequences identified up to six psbA copies per genome, with always a single gene encoding D1: 1. In phylogenetic analyses, marine Synechococcus genes encoding D1: 1 clustered together, while the genes encoding D1: 2 grouped by genome into subclusters. Moreover, examination of the genomic environment of psbA genes suggests that the D1: 2 genes are hotspots for DNA recombination. Collectively, our observations suggest that while all psbA genes follow a concerted evolution within each genome, D1: 2 coding genes are subject to intragenome homogenization most probably mediated by gene conversion.

Phytoplankton function and acclimation are driven by catalytic protein complexes that mediate key physiological transformations, including generation of photosynthetic ATP and reductant, and carbon and nitrogen fixation. Quantitation of capacities for these processes allows estimation of rates for key ecosystem processes, and identification of factors limiting primary productivity. We herein present molar quantitations of PSI, PSII, ATP synthase, RuBisCO and the Fe protein of nitrogenase of Trichodesmium collected from the Gulf of Mexico, in comparison to determinations for a range of cyanobacteria growing in culture. Using these measurements, estimates were generated for Trichodesmium capacities for carbon fixation of 1–3.4 g C g chl a −1 h−1 and nitrogen fixation of 0.06–0.17 g N g chl a −1 h−1, with diel variations in capacities. ATP synthase levels show that ATP synthesis capacity is sufficient to support these levels of carbon and nitrogen fixation, and that ATP synthase levels change over the day in accordance with the ATP demands of nitrogenase and RuBisCO activity. Levels of measured complexes indicate that Trichodesmium manifests n-type diel light acclimation through rapid changes in RuBisCO:PSII, supported by significant investment of cellular nitrogen. The plasticity in the levels and stoichiometry of these core complexes show that changes in the abundance of core protein complexes are an important component of acclimation and regulation of metabolic function by Trichodesmium populations.

Gloeobacter violaceus PCC 7421 is a slow-growing cyanobacterium which lacks thylakoid membranes, but whose five-membered psbA gene family encodes three isoform variants of the PsbA (D1) reaction center protein of Photosystem II. Under standard culture conditions Gloeobacter exhibits photosystem II electron transport, but several clear modifications in the redox potential of key cofactors bound by the PsbA protein are manifested in the flash-fluorescence characteristics. In other cyanobacteria dynamic expression of multiple psbA genes and turnover of PsbA isoforms is critical to counter excitation stress. We found that each of Gloeobacter's five psbA genes is expressed, with transcript abundances spanning 4.5 orders of magnitude. psbAI (glr2322) and psbAII (glr0779), encoding identical PsbA:2 form proteins, are constitutively expressed and dominate the psbA transcript pool under control conditions. psbAIII (gll3144) was strongly induced under photoinhibitory high irradiance stress, thereby contributing to a large increase in the psbA transcript pool that allowed cells to maintain their PsbA protein pools and then recover from irradiance stress, within one cellular generation. In contrast, under comparable photoinhibition provoked by UVB the cells were unable to maintain their psbA transcript and PsbA protein pools, and showed limited subsequent recovery. psbAIV (glr1706) and psbAV (glr2656), encoding two divergent PsbA isoforms, showed consistent trace expression but were never quantitatively significant contributors to the psbA transcript pool.

Spartina alterniflora Loisel. is a dominant primary producer in salt marsh intertidal zones along the Bay of Fundy of New Brunswick and Nova Scotia, Canada, where it has a 5-month growing season from May to September. We quantified key subunits of the S. alterniflora photosynthetic system (chlorophyll, PsbA, PsaC, AtpB, RbcL) over a seasonal developmental progression to determine resource allocations and estimate capacities for key subprocesses of photosynthesis catalyzed by light-harvesting antennae, photosystems I and II (PSI and PSII, respectively), ATP (adenosine triphosphate) synthase, and RuBisCO (ribulose bisphosphate carboxylase–oxygenase). Early in the season, all of the macromolecules peaked in concentration around day 159, but then declined as leaves elongated. Later in the season, just after flowering, chlorophyll b, PsbA, PsaC, and AtpB peaked again before declining at the end of the season. RbcL, however, did not exhibit a second late-season peak, but continually declined from the early-season peak. Estimates of metabolic flux per PSII and per RuBisCO were closely parallel until late in the season, indicating coordinated regulation of catalytic turnover per complex. Early in the season, carbon fixation per RuBisCO peaked near the expected turnover rate (kcat) for a C4 plant, but then declined. The effective absorbance cross sections of PSII varied seasonally, reflecting significant regulation of antenna performance across the season.

Cyanobacteria show high metabolic plasticity by re-allocating macromolecular resources in response to variations in both environmental inorganic carbon (Ci) and light. We grew cultures of the picoplanktonic cyanobacterium Synechococcus elongatus Nageli across a 50-fold range of growth irradiance at either a dissolved [Ci]

Cyanobacteria cope with UVB induced photoinhibition of Photosystem II by regulating multiple psbA genes to boost the expression of D1 protein (in Synechocystis sp. PCC6803), or to exchange the constitutive D1:1 protein to an alternate D1:2 isoform (in Synechococcus sp. PCC7942). To define more general patterns of cyanobacterial psbA expression, we applied moderately photoinhibitory UVB to Anabaena sp. PCC7120 and tracked the expression of its five psbA genes. psbAI, encoding a D1:1 protein isoform characterized by a Gln130, represented the majority of the psbA transcript pool under control conditions. psbAI transcripts decreased upon UVB treatment but the total psbA transcript pool increased 3.5 fold within 90 min as a result of sharply increased psbAII, psbAIV and psbAIII transcripts encoding an alternate D1:2 protein isoform characterized by Glu130, similar to that of Synechococcus. Upon UVB treatment the relaxation of flash induced chlorophyll fluorescence showed a characteristic acceleration of a decay phase likely associated with the exchange from the D1:1 protein isoform encoded by psbAI to the alternate D1:2 isoform encoded by psbAIV, psbAII and psbAIII. Throughout the UVB treatment the divergent psbA0 made only a trace contribution to the total psbA transcript pool. This suggests a similarity to the divergent psbAI gene from Synechocystis, whose natural expression we demonstrate for the first time at a trace level similar to psbA0 in Anabaena. These trace-expressed psbA genes in two different cyanobacteria raise questions concerning the functions of these divergent genes.

Photosynthetic efficiency in cyanobacteria is improved under conditions of inorganic carbon (Ci) limitation by the induction of a CO2-concentrating mechanism (CCM) that elevates the CO2 concentration around the primary carboxylase ribulose-1,5-bisphosphate carboxylase–oxygenase (Rubisco) in the carboxysome. The molecular details of low-Ci sensing in relation to regulation of CCM induction in cyanobacteria are not presently known. We have applied a quantitative reverse transcription – polymerase chain reaction technique to monitor the abundance of key CCM-related transcripts in Synechococcus elongatus under a variety of experimental conditions with the aim of probing the conditions required for CCM induction. Despite preliminary evidence for strong induction of cmpA, sbtA, and chpY transcripts in the dark under low Ci in Synechococcus elongatus, subsequent experiments in which contaminating levels of room light during harvest were eliminated demonstrated that light is required for induction of these transcripts. However, the requirement for light for significant accumulation of CCM-related transcripts was very modest and well below the light level required to induce measurable net photosynthetic electron transport. Brief treatments with weak monochromatic light under low Ci were sufficient to cause significant accumulation of transcripts in Synechococcus elongatus relative to cells held in continuous darkness.Key words: CO2-concentrating mechanism, cyanobacteria, photosynthesis.

Acclimation of cyanobacteria to ambient fluctuations in inorganic carbon (Ci) and temperature requires reorganization of the major protein complexes involved in photosynthesis. We grew cultures of the picoplanktonic cyanobacterium Synechococcus elongatus Naegeli across most of its range of tolerable temperatures from 23 to 35° C at both low (