Study on photosynthetic responses and chlorophyll fluorescence in Rhizophora mucronata seedlings under shade regimes

The photosynthetic performance of mangrove Rhizophora mucronata seedlings grown under seasonally full light (HL), 50 % shade (ML), and 80 % shade (LL) conditions was characterized by gas exchange, and chlorophyll fluorescence. The carboxylation efficiency significantly affected the seasonal change of the photosynthetic capacity. Temperature and light might have synergic effect on the carboxylation efficiency. The photosynthetic rate (PN) of R. mucronata seedlings under shade regimes, however, could not be attributed to variability in chlorophyll, C _i , ΦPSII, ETR or qP values but more to differences in carboxylation efficiency, g _max, and E _max. HL and ML plants had higher PN, g _s and E than the LL ones. Nevertheless, LL leaves exhibited low photoinhibition susceptibility. The high non-photochemical quenching in HL leaves may show that applied light intensity probably exceeded the photosynthetic capability. The findings indicate that ML treatments provided the best condition to obtain such carbon fixation capacity.     

Photosynthetic acclimation to different light levels in the brown marine macroalga, <Emphasis Type="Italic">Hizikia fusiformis</Emphasis> (Sargassaceae,Phaeophyta)

Hizikia fusiformis thalli experience dynamic incident light conditions during the period of growth. The present study was designed to examine how changing photon irradiance affects the photosynthesis both in the short and long terms by culturing H. fusiformis under three different light levels: 35 μmol photons m^-2 s^-1 (low light, LL), 85 μmol photons m^-2 s^-1 (intermediate light, IL), and 165 μmol photons m^-2 s^-1 (high light, HL). A similar relative growth rate was observed between IL- and HL-grown algae, but the growth rate was significantly reduced in LL-grown algae. The photosynthetic rates (P _n) measured at their respective growth light levels were found to be lowest in the thalli grown at LL and highest at HL. However, LL-grown algae exhibited much higher P _n in comparison with IL- and the HL-grown thalli at the same measuring photosynthetic photon flux density, indicating the photosynthetic acclimation to low growth light in H. fusiformis. The photosynthesis–light curves showed that LL-grown algae had a highest light-saturating maximum P _n (P _max) in comparison with IL- or HL-grown algae when the photosynthetic rates were expressed on the biomass basis. However, P _max was highest in HL-grown algae compared to IL- or LL-grown algae when the rates were normalized to chlorophyll a. The photosynthesis–inorganic carbon (Ci) response curves were also significantly affected by the growth light conditions. The highest value of apparent photosynthetic conductance occurred in LL-grown algae while the lowest value in HL-grown algae. Additionally, the activity of external carbonic anhydrase (CA) tended to increase while the total CA activity inclined to decrease in H. fusiformis thalli when the growth light level altered from 35 to 165 μmol photons per square meter per second. The external CA inhibitors showed a higher inhibition in HL-grown algae compared with LL-grown algae. It was proposed that photosynthetic acclimation to low light condition in H. fusiformis was achieved through an increase in the number of reaction centers and increased capacities of electron transport and of Ci transport within cells. The ability of photosynthetic acclimation to low light confers H. fusiformis thalli to overcome the environmental low light condition as a result of the attenuation of seawater or self-shading through enhancing its photosynthetic performance and carbon assimilation necessary for growth.     

NITROGEN FIXATION,HYDROGEN CYCLING,AND ELECTRON TRANSPORT KINETICS IN TRICHODESMIUM ERYTHRAEUM (CYANOBACTERIA) STRAIN IMS1011

This study describes the relationships between dinitrogen (N₂) fixation, dihydrogen (H₂) production, and electron transport associated with photosynthesis and respiration in the marine cyanobacterium Trichodesmium erythraeum Ehrenb. strain IMS101. The ratio of H₂ produced:N₂ fixed (H₂:N₂) was controlled by the light intensity and by the light spectral composition and was affected by the growth irradiance level. For Trichodesmium cells grown at 50 μmol photons · m^?2 · s^?1, the rate of N₂ fixation, as measured by acetylene reduction, saturated at light intensities of 200 μmol photons · m^?2 · s^?1. In contrast, net H₂ production continued to increase with light levels up to 1,000 μmol photons · m^?2 · s^?1. The H₂:N₂ ratios increased monotonically with irradiance, and the variable fluorescence measured using a fast repetition rate fluorometer (FRRF) revealed that this increase was accompanied by a progressive reduction of the plastoquinone (PQ) pool. Additions of 2,5‐dibromo‐3‐methyl‐6‐isopropyl‐p‐benzoquinone (DBMIB), an inhibitor of electron transport from PQ pool to PSI, diminished both N₂ fixation and net H₂ production, while the H₂:N₂ ratio increased with increasing level of PQ pool reduction. In the presence of 3‐(3,4‐dichlorophenyl)‐1,1‐dimethylurea (DCMU), nitrogenase activity declined but could be prolonged by increasing the light intensity and by removing the oxygen supply. These results on the coupling of N₂ fixation and H₂ cycling in Trichodesmium indicate how light intensity and light spectral quality of the open ocean can influence the H₂:N₂ ratio and modulate net H₂ production.     

Modeling the Dynamic Regulation of Nitrogen Fixation in the Cyanobacterium Trichodesmium sp.

A physiological, unbalanced model is presented that explicitly describes growth of the marine cyanobacterium Trichodesmium sp. at the expense of N₂ (diazotrophy). The model involves the dynamics of intracellular reserves of carbon and nitrogen and allows the uncoupling of the metabolism of these elements. The results show the transient dynamics of N₂ fixation when combined nitrogen (NO₃⁻, NH₄⁺) is available and the increased rate of N₂ fixation when combined nitrogen is insufficient to cover the demand. The daily N₂ fixation pattern that emerges from the model agrees with measurements of rates of nitrogenase activity in laboratory cultures of Trichodesmium sp. Model simulations explored the influence of irradiance levels and the length of the light period on fixation activity and cellular carbon and nitrogen stoichiometry. Changes in the cellular C/N ratio resulted from allocations of carbon to different cell compartments as demanded by the growth of the organism. The model shows that carbon availability is a simple and efficient mechanism to regulate the balance of carbon and nitrogen fixed (C/N ratio) in filaments of cells. The lowest C/N ratios were obtained when the light regime closely matched nitrogenase dynamics.     

Combined Effects of CO2 and Light on the N2-Fixing Cyanobacterium Trichodesmium IMS101: A Mechanistic View

The marine diazotrophic cyanobacterium Trichodesmium responds to elevated atmospheric CO₂ partial pressure (pCO₂) with higher N₂ fixation and growth rates. To unveil the underlying mechanisms, we examined the combined influence of pCO₂ (150 and 900 μatm) and light (50 and 200 μmol photons m⁻² s⁻¹) on Trichodesmium IMS101. We expand on a complementary study that demonstrated that while elevated pCO₂ enhanced N₂ fixation and growth, oxygen evolution and carbon fixation increased mainly as a response to high light. Here, we investigated changes in the photosynthetic fluorescence parameters of photosystem II, in ratios of the photosynthetic units (photosystem I:photosystem II), and in the pool sizes of key proteins involved in the fixation of carbon and nitrogen as well as their subsequent assimilation. We show that the combined elevation in pCO₂ and light controlled the operation of the CO₂-concentrating mechanism and enhanced protein activity without increasing their pool size. Moreover, elevated pCO₂ and high light decreased the amounts of several key proteins (NifH, PsbA, and PsaC), while amounts of AtpB and RbcL did not significantly change. Reduced investment in protein biosynthesis, without notably changing photosynthetic fluxes, could free up energy that can be reallocated to increase N₂ fixation and growth at elevated pCO₂ and light. We suggest that changes in the redox state of the photosynthetic electron transport chain and posttranslational regulation of key proteins mediate the high flexibility in resources and energy allocation in Trichodesmium. This strategy should enable Trichodesmium to flourish in future surface oceans characterized by elevated pCO₂, higher temperatures, and high light.The marine filamentous N₂-fixing (diazotrophic) cyanobacteria Trichodesmium spp. bloom extensively in the oligotrophic subtropical and tropical oceans (Carpenter and Capone, 2008). Trichodesmium contributes 25% to 50% of the estimated rates of N₂ fixation in these areas, where the new nitrogen inputs stimulate carbon and nitrogen cycling (Capone and Subramaniam, 2005; Mahaffey et al., 2005). The increases in atmospheric CO₂ partial pressure (pCO₂) and the subsequent impacts on ocean acidification are predicted to influence diazotrophs and specifically Trichodesmium.The reported sensitivity of Trichodesmium to changes in pCO₂ prompted further investigation into the cellular responses and underlying mechanisms, specifically when combined with other environmental parameters such as temperature, nutrient availability, and light. Elevated pCO₂ significantly increased growth and N₂ fixation rates of Trichodesmium cultures (Barcelos é Ramos et al., 2007; Hutchins et al., 2007; Levitan et al., 2007, 2010). The physiological response was also characterized by changes in inorganic carbon acquisition, limited flexibility of carbon-nitrogen ratios, and conservation of photosynthetic activities with increased pCO₂. These manifestations suggested that ATP and reductants [ferredoxin, NAD(P)H] are reallocated in the cells (Levitan et al., 2007, 2010; Kranz et al., 2009, 2010).In Trichodesmium, as in all cyanobacteria, the metabolic pathways of respiration and photosynthesis share several cellular complexes/proteins such as the plastoquinone (PQ) pool, succinate dehydrogenase, and ferredoxin (Fig. 1; Kana, 1993; Bergman et al., 1997; Lin et al., 1998). Energetic currencies [reduced ferredoxin, ATP, NAD(P)H] are also shared and can be allocated and utilized according to cellular requirements. N₂ fixation by nitrogenase and the subsequent assimilation of NH₄⁺ by Gln synthetase requires carbon skeletons from the tricarboxylic acid reactions. Moreover, linear and pseudocyclic photosynthesis can also generate additional ATP and reductants essential for N₂ fixation (Fig. 1; Berman-Frank et al., 2001).Open in a separate windowFigure 1.Schematic representation of major cellular complexes involved in energy flow [electron, ATP, NAD(P)H, carbon skeletons] in Trichodesmium IMS101. Dashed arrows represent movement direction of electrons, and solid arrows represent directions of protons, ATP, and NAD(P)H. Measured protein subunits are represented by gray diamonds. See Kranz et al. (2010) for measurements of O₂ evolution, inorganic carbon fixation, and fluxes of N₂ fixation.To understand the regulation of these metabolic pathways in Trichodesmium under varying pCO₂ levels and light intensities, we designed an experiment to characterize changes in the fluxes of carbon, nitrogen, and oxygen (O₂), related protein pool sizes, and variable fluorescence parameters of PSII. Elevated atmospheric pCO₂ combined with enhanced sea surface temperatures are forecast to stabilize thermal stratification, resulting in a shallower, more acidified, upper mixed layer characterized by higher mean light intensities (Doney, 2006). Thus, Trichodesmium IMS101 cultures were acclimated to past and future pCO₂ levels (150 and 900 μatm) at low and high light (50 and 200 μmol photons m⁻² s⁻¹).In the first part of this combined report (Kranz et al., 2010), we examined the physiological responses to the different acclimation conditions. The combination of elevated pCO₂ and light enhanced the production of particulate organic carbon and nitrogen (270% and 390% increase, respectively) as well as growth rates (180% increase; percentages are calculated from Kranz et al., 2010). Generally, the pCO₂-dependent stimulation was higher in cultures acclimated to low light. The pCO₂ effect was also reflected in other measured physiological parameters, particularly the diel patterns of N₂ fixation and the integrated N₂ fixation rates during the day, which increased approximately 30-fold between the low-pCO₂/low-light and the high-pCO₂/high-light acclimations (Kranz et al., 2010). While at high light, elevated pCO₂ extended the period of high N₂ fixation, which lasted from 5 h after the onset of light throughout the end of the photoperiod, the high-pCO₂ contribution to the integrated N₂ fixation was more significant at low light (Kranz et al., 2010). Light, but not pCO₂, influenced gross photosynthesis as measured by PSII O₂ evolution, which increased by approximately 250% in high-light-acclimated cultures. To supply the Calvin cycle with sufficient CO₂, Trichodesmium possesses a CO₂-concentrating mechanism mainly based on HCO₃⁻ uptake (Kranz et al., 2009, 2010). When Trichodesmium was acclimated to elevated pCO₂ (900 μatm), a decline in the cellular affinity to dissolved inorganic carbon was observed (Kranz et al., 2009), while the specific uptake of CO₂ showed a 9-fold increase between the low-pCO₂/low-light and the high-pCO₂/high-light acclimations (Kranz et al., 2010).Proteins are fundamental cellular components that influence the underlying mechanisms subsequently reflected in the cells’ physiology. In this study, we extend the experimental results presented by Kranz et al. (2010) by examining the influence of pCO₂ at different light regimes on the photosynthetic fluorescence parameters of PSII and on the pool sizes of key proteins involved in carbon and nitrogen fixation and their subsequent assimilation processes.     

INTERACTIVE EFFECTS OF IRRADIANCE AND CO2 ON CO2 FIXATION AND N2 FIXATION IN THE DIAZOTROPH TRICHODESMIUM ERYTHRAEUM (CYANOBACTERIA)1

The diazotrophic cyanobacteria Trichodesmium spp. contribute approximately half of the known marine dinitrogen (N₂) fixation. Rapidly changing environmental factors such as the rising atmospheric partial pressure of carbon dioxide (pCO₂) and shallower mixed layers (higher light intensities) are likely to affect N₂‐fixation rates in the future ocean. Several studies have documented that N₂ fixation in laboratory cultures of T. erythraeum increased when pCO₂ was doubled from present‐day atmospheric concentrations (～380 ppm) to projected future levels (～750 ppm). We examined the interactive effects of light and pCO₂ on two strains of T. erythraeum Ehrenb. (GBRTRLI101 and IMS101) in laboratory semicontinuous cultures. Elevated pCO₂ stimulated gross N₂‐fixation rates in cultures growing at 38 μmol quanta · m^?2 · s^?1 (GBRTRLI101 and IMS101) and 100 μmol quanta · m^?2 · s^?1 (IMS101), but this effect was reduced in both strains growing at 220 μmol quanta · m^?2 · s^?1. Conversely, CO₂‐fixation rates increased significantly (P < 0.05) in response to high pCO₂ under mid‐ and high irradiances only. These data imply that the stimulatory effect of elevated pCO₂ on CO₂ fixation and N₂ fixation by T. erythraeum is correlated with light. The ratio of gross:net N₂ fixation was also correlated with light and trichome length in IMS101. Our study suggests that elevated pCO₂ may have a strong positive effect on Trichodesmium gross N₂ fixation in intermediate and bottom layers of the euphotic zone, but perhaps not in light‐saturated surface layers. Climate change models must consider the interactive effects of multiple environmental variables on phytoplankton and the biogeochemical cycles they mediate.     

BACTERIAL ASSOCIATIONS WITH MARINE OSCILLATORIA SP. (TRICHODESMIUM SP.) POPULATIONS: ECOPHYSIOLOGICAL IMPLICATIONS1

On three separate occasions we investigated morphological and physiological aspects of bacterial associations with planktonic aggregates of the ubiquitous marine N₂ fixing cyanobacterium Trichodesmium sp. Close associations generally characterized Trichodesmium blooms; associations were present during day- and night-time. Colonization by both rod-shaped and filamentous heterotrophic bacteria occurred on Trichodesmiun aggregates actively fixing N₂ (acetylene reduction). Scanning electron and optical microscopy showed bacteria located both around and within aggregates. Microautoradiography demonstrated that associated bacteria largely mediated utilization of trace additions of ³H-labeled carbohydrates (fructose, glucose, mannitol) and amino acids, whereas Trichodesmium utilized amino acids only. Oxygen measurements using microelectrodes revealed high localized oxygen consumption among aggregates, with rapid (within a minute) changes from supersaturated to subsaturated oxygen following the transition from photosynthetic illuminated to dark periods. Stab culturing techniques confirmed the presence of heterotrophic N₂ fixers among aggregate-associated bacteria. Parallel deployment of oxygen microelectrodes, the tetrazolium salt 2,3,5 triphenyl tetrazolium chloride (TTC) and acetylene reduction assays demonstrated microaerophilic requirements for expression of nitrogenase activity among cultured bacteria. Trichodesmium aggregates are characterized by dynamic nutrient and oxygen regimes, which promote and maintain simultaneous and contiguous oxygenic photosynthesis and N₂ fixation. In part, the above-mentioned consortial interactions with a variety of heterotrophic bacteria facilitate Trichodesmium biomass production and bloom formation in nitrogen depleted, oligotrophic tropical/subtropical waters.     

Combined Effects of CO2 and Light on the N2-Fixing Cyanobacterium Trichodesmium IMS101: Physiological Responses

Recent studies on the diazotrophic cyanobacterium Trichodesmium erythraeum (IMS101) showed that increasing CO₂ partial pressure (pCO₂) enhances N₂ fixation and growth. Significant uncertainties remain as to the degree of the sensitivity to pCO₂, its modification by other environmental factors, and underlying processes causing these responses. To address these questions, we examined the responses of Trichodesmium IMS101 grown under a matrix of low and high levels of pCO₂ (150 and 900 μatm) and irradiance (50 and 200 μmol photons m⁻² s⁻¹). Growth rates as well as cellular carbon and nitrogen contents increased with increasing pCO₂ and light levels in the cultures. The pCO₂-dependent stimulation in organic carbon and nitrogen production was highest under low light. High pCO₂ stimulated rates of N₂ fixation and prolonged the duration, while high light affected maximum rates only. Gross photosynthesis increased with light but did not change with pCO₂. HCO₃⁻ was identified as the predominant carbon source taken up in all treatments. Inorganic carbon uptake increased with light, but only gross CO₂ uptake was enhanced under high pCO₂. A comparison between carbon fluxes in vivo and those derived from ¹³C fractionation indicates high internal carbon cycling, especially in the low-pCO₂ treatment under high light. Light-dependent oxygen uptake was only detected under low pCO₂ combined with high light or when low-light-acclimated cells were exposed to high light, indicating that the Mehler reaction functions also as a photoprotective mechanism in Trichodesmium. Our data confirm the pronounced pCO₂ effect on N₂ fixation and growth in Trichodesmium and further show a strong modulation of these effects by light intensity. We attribute these responses to changes in the allocation of photosynthetic energy between carbon acquisition and the assimilation of carbon and nitrogen under elevated pCO₂. These findings are supported by a complementary study looking at photosynthetic fluorescence parameters of photosystem II, photosynthetic unit stoichiometry (photosystem I:photosystem II), and pool sizes of key proteins in carbon and nitrogen acquisition.Human-induced climate change will significantly alter the marine environment within the next century and beyond. Future scenarios predict an increase from currently approximately 380 to about 750 to 1,000 μatm CO₂ partial pressure (pCO₂) in the atmosphere until the end of this century (Raven et al., 2005; Raupach et al., 2007). As the ocean takes up this anthropogenic CO₂, dissolved inorganic carbon (DIC) in the surface ocean increases while the pH decreases (Wolf-Gladrow et al., 1999). Rising global temperatures will increase surface ocean stratification, which may affect the light regime in the upper mixed layer as well as nutrient input from deeper waters (Doney, 2006). Uncertainties remain regarding both the magnitude of the physicochemical changes and the biological responses of organisms, including species and populations of the oceanic primary producers at the basis of the food webs.In view of potential ecological implications and feedbacks on climate, several studies have examined pCO₂ sensitivity in phytoplankton key species (Burkhardt and Riebesell, 1997; Riebesell et al., 2000; Rost et al., 2003; Tortell et al., 2008). Pronounced responses to elevated pCO₂ were observed in N₂-fixing cyanobacteria (Barcelos é Ramos et al., 2007; Hutchins et al., 2007; Levitan et al., 2007; Fu et al., 2008; Kranz et al., 2009), which play a vital role in marine ecosystems by providing a new source of biologically available nitrogen species to otherwise nitrogen-limited regions. Recent studies focused on the impact of different environmental factors on the filamentous Trichodesmium species, which is known for high abundance and the formation of massive blooms in tropical and subtropical areas (Capone et al., 2005; Mahaffey et al., 2005). Higher pCO₂ levels stimulated growth rates, biomass production, and N₂ fixation (Hutchins et al., 2007; Levitan et al., 2007; Kranz et al., 2009) and affected inorganic carbon acquisition of the cells (Kranz et al., 2009). While elevated sea surface temperatures are predicted to shift the spatial distribution of Trichodesmium species toward higher latitudes (Breitbarth et al., 2007), the combined effects of pCO₂ and temperature may favor this species and extend its niche even farther (Hutchins et al., 2007; Levitan et al., 2010a). An increase in the average light intensity, caused by the predicted shoaling of the upper mixed layer, may further stimulate photosynthesis and thus growth and N₂ fixation of Trichodesmium (Breitbarth et al., 2008). To our knowledge, the combined effects of light and pCO₂ have not been studied yet, although these environmental factors are likely to influence photosynthesis and other key processes in Trichodesmium.To understand the responses of an organism to changes in environmental conditions, metabolic processes must be studied. In Trichodesmium, photosynthetically generated energy (ATP and NADPH) is primarily used for the fixation of CO₂ in the Calvin-Benson cycle. A large proportion of this energy, however, is also required for the process of N₂ fixation via nitrogenase and for the operation of a CO₂-concentrating mechanism (CCM). The latter involves active uptake of inorganic carbon, which functions to increase the rate of carboxylation reaction mediated by Rubisco. This ancient and highly conserved enzyme is characterized by low affinities for its substrate CO₂ and a susceptibility to a competing reaction with oxygen (O₂) as substrate (Badger et al., 1998); the latter initiates photorespiration. As cyanobacterial Rubisco possesses one of the lowest CO₂ affinities among phytoplankton (Badger et al., 1998), a considerable amount of resources have to be invested to achieve sufficient rates of carbon fixation and to avoid photorespiration. A first step toward a mechanistic understanding of responses in Trichodesmium has been taken by Levitan et al. (2007), focusing on pCO₂ dependency of nitrogenase activity and photosynthesis. Subsequently, Kranz et al. (2009) described variations in CCM efficiency with pCO₂ and suggested that the observed plasticity in CCM regulation allowed energy reallocation under high pCO₂, which may explain the observed pCO₂-dependent changes in nitrogenase activity, growth, and elemental composition (Barcelos é Ramos et al., 2007; Hutchins et al., 2007; Levitan et al., 2007).In this study, we measured growth responses as well as metabolic key processes in Trichodesmium erythraeum (IMS101) under environmental conditions that likely alter the energy budget and/or energy allocation of the cell. Cultures were acclimated to a matrix of low and high pCO₂ (150 and 900 μatm) at two different light intensities (50 and 200 μmol photons m⁻² s⁻¹). For each of the four treatments, changes in growth rates, elemental ratios, and the accumulation of particulate carbon and nitrogen were measured. Metabolic processes (gross photosynthesis, CCM activity, and O₂ uptake) were obtained by means of membrane-inlet mass spectrometry (MIMS), while N₂ fixation was detected by gas chromatography. As these processes may vary over the diurnal cycle in Trichodesmium (Berman-Frank et al., 2001; Kranz et al., 2009), measurements were performed in the morning and around midday. The results on metabolic processes were accompanied by measurements of the fluorescence of PSII, ratios of the photosynthetic units (PSI:PSII), and pool sizes of key proteins involved in carbon and nitrogen fixation as well as assimilation (Levitan et al., 2010b).     

Acclimation to low light by C4 maize: implications for bundle sheath leakiness

C4 plants have a biochemical carbon concentrating mechanism (CCM) that increases CO₂ concentration around ribulose bisphosphate carboxylase oxygenase (Rubisco) in the bundle sheath (BS). Under limiting light, the activity of the CCM generally decreases, causing an increase in leakiness, (Φ), the ratio of CO₂ retrodiffusing from the BS relative to C4 carboxylation processes. Maize plants were grown under high and low light regimes (respectively HL, 600 versus LL, 100 μE m^?2 s^?1). Short‐term acclimation of Φ was compared from isotopic discrimination (Δ), gas exchange and photochemistry. Direct measurement of respiration in the light, and ATP production rate (J_ATP), allowed us use a novel approach to derive Φ, compared with the conventional fitting of measured and predicted Δ. HL grown plants responded to decreasing light intensities with the well‐documented increase in Φ. Conversely, LL plants showed a constant Φ, which has not been observed previously. We explain the pattern by two contrasting acclimation strategies: HL plants maintained a high CCM activity at LL, resulting in high CO₂ overcycling and increased Φ; LL plants acclimated by down‐regulating the CCM, effectively optimizing scarce ATP supply. This surprising plasticity may limit the impact of Φ‐dependent carbon losses in leaves becoming shaded within developing canopies.     

Response of the diatom Phaeodactylum tricornutum to photooxidative stress resulting from high light exposure

The response of microalgae to photooxidative stress resulting from high light exposure is a well-studied phenomenon. However, direct analyses of photosystem II (PSII) D1 protein (the main target of photoinhibition) in diatoms are scarce. In this study, the response of the diatom model species Phaeodactylum tricornutum to short-term exposure to high light was examined and the levels of D1 protein determined immunochemically. Low light (LL) acclimated cells (40 μmol photons m(-2) s(-1)) subjected to high light (HL, 1,250 μmol photons m(-2) s(-1)) showed rapid induction of non-photochemical quenching (NPQ) and ca. 20-fold increase in diatoxanthin (DT) concentration. This resulted from the conversion of diadinoxanthin (DD) to DT through the activation of the DD-cycle. D1 protein levels under LL decreased about 30% after 1 h of the addition of lincomycin (LINC), a chloroplast protein synthesis inhibitor, showing significant D1 degradation and repair under low irradiance. Exposure to HL lead to a 3.2-fold increase in D1 degradation rate, whereas average D1 repair rate was 1.3-x higher under HL than LL, leading to decreased levels of D1 protein under HL. There were significant effects of both HL and LINC on P. tricornutum maximum quantum yield of PSII (F(v)/F(m)), showing a reduction of active PSII reaction centres. Partial recovery of F(v)/F(m) in the dark demonstrates the photosynthetic resilience of this diatom to changes in the light regime. P. tricornutum showed high allocation of total protein to D1 and an active D1-repair cycle to limit photoinhibition.     

Acclimation of shade-tolerant and light-resistant <Emphasis Type="Italic">Tradescantia</Emphasis> species to growth light: chlorophyll <Emphasis Type="Italic">a</Emphasis> fluorescence,electron transport,and xanthophyll content

In this study, we have compared the photosynthetic characteristics of two contrasting species of Tradescantia plants, T. fluminensis (shade-tolerant species), and T. sillamontana (light-resistant species), grown under the low light (LL, 50–125 µmol photons m^?2 s^?1) or high light (HL, 875–1000 µmol photons m^?2 s^?1) conditions during their entire growth period. For monitoring the functional state of photosynthetic apparatus (PSA), we measured chlorophyll (Chl) a emission fluorescence spectra and kinetics of light-induced changes in the heights of fluorescence peaks at 685 and 740 nm (F ₆₈₅ and F ₇₄₀). We also compared the light-induced oxidation of P₇₀₀ and assayed the composition of carotenoids in Tradescantia leaves grown under the LL and HL conditions. The analyses of slow induction of Chl a fluorescence (SIF) uncovered different traits in the LL- and HL-grown plants of ecologically contrasting Tradescantia species, which may have potential ecophysiological significance with respect to their tolerance to HL stress. The fluorometry and EPR studies of induction events in chloroplasts in situ demonstrated that acclimation of both Tradescantia species to HL conditions promoted faster responses of their PSA as compared to LL-grown plants. Acclimation of both species to HL also caused marked changes in the leaf anatomy and carotenoid composition (an increase in Violaxanthin?+?Antheraxantin?+?Zeaxanthin and Lutein pools), suggesting enhanced photoprotective capacity of the carotenoids in the plants grown in nature under high irradiance. Collectively, the results of the present work suggest that the mechanisms of long-term PSA photoprotection in Tradescantia are based predominantly on the light-induced remodeling of pigment-protein complexes in chloroplasts.     

LIGHT‐INDUCED RESPONSES OF OXYGEN PHOTOREDUCTION,REACTIVE OXYGEN SPECIES PRODUCTION AND SCAVENGING IN TWO DIATOM SPECIES1

Diatoms are frequently exposed to high light (HL) levels, which can result in photoinhibition and damage to PSII. Many microalgae can photoreduce oxygen using the Mehler reaction driven by PSI, which could protect PSII. The ability of Nitzschia epithemioides Grunow and Thalassiosira pseudonana Hasle et Heimdal grown at 50 and 300 μmol photons · m^?2 · s^?1 to photoreduce oxygen was examined by mass spectrometric measurements of ¹⁸O₂. Both species exhibited significant rates of oxygen photoreduction at saturating light levels, with cells grown in HL exhibiting higher rates. HL‐grown T. pseudonana had maximum rates of oxygen photoreduction five times greater than N. epithemoides, with 49% of electrons transported through PSII being used to reduce oxygen. Exposure to excess light (1,000 μmol photons · m^?2 · s^?1) produced similar decreases in the operating quantum efficiency of PSII (F_q′/F_m′) of low light (LL)‐ and HL‐grown N. epithemoides, whereas HL‐grown T. pseudonana exhibited much smaller decreases in F_q′/F_m′ than LL‐grown cells. HL‐grown T. pseudonana and N. epithemioides exhibited greater superoxide and hydrogen peroxide production, higher activities (in T. pseudonana) of superoxide dismutase (SOD) and ascorbate peroxidase (APX), and increased expression of three SOD‐ and one APX‐encoding genes after 60 min of excess light compared to LL‐grown cells. These responses provide a mechanism that contributes to the photoprotection of PSII against photodamage.     

Importance of AOX pathway in optimizing photosynthesis under high light stress: role of pyruvate and malate in activating AOX

The present study shows the importance of alternative oxidase (AOX) pathway in optimizing photosynthesis under high light (HL). The responses of photosynthesis and respiration were monitored as O₂ evolution and O₂ uptake in mesophyll protoplasts of pea pre‐incubated under different light intensities. Under HL (3000 µmol m^?2 s^?1), mesophyll protoplasts showed remarkable decrease in the rates of NaHCO₃‐dependent O₂ evolution (indicator of photosynthetic carbon assimilation), while decrease in the rates of respiratory O₂ uptake were marginal. While the capacity of AOX pathway increased significantly by two fold under HL, the capacity of cytochrome oxidase (COX) pathway decreased by >50% compared with capacities under darkness and normal light (NL). Further, the total cellular levels of pyruvate and malate, which are assimilatory products of active photosynthesis and stimulators of AOX activity, were increased remarkably parallel to the increase in AOX protein under HL. Upon restriction of AOX pathway using salicylhydroxamic acid (SHAM), the observed decrease in NaHCO₃‐dependent O₂ evolution or p‐benzoquinone (BQ)‐dependent O₂ evolution [indicator of photosystem II (PSII) activity] and the increase in total cellular levels of pyruvate and malate were further aggravated/promoted under HL. The significance of raised malate and pyruvate levels in activation of AOX protein/AOX pathway, which in turn play an important role in dissipating excess chloroplastic reducing equivalents and sustenance of photosynthetic carbon assimilation to balance the effects of HL stress on photosynthesis, was depicted as a model.     

Growth,N2 fixation and photosynthesis in a cyanobacterium,Trichodesmium sp., under Fe stress

Trichodesmium sp., isolated from the Great Barrier Reef lagoon, was cultured in artificial seawater media containing a range of Fe concentration. Fe additions stimulated growth, N₂ fixation, cellular chlorophyll a content, light-saturated chlorophyll a-specific gross photosynthetic capacity (P_m ^chla) and the dark respiration rate (R_d ^chla). Cell yields only doubled for 9 nM Fe relative to zero added Fe, whereas N₂ fixation increased 11-fold considerably for 450 nM Fe. The results suggest that N₂ fixation of Trichodesmium is more sensitive to Fe limitation than are the cell yields.     

DIAZOTROPHIC GROWTH OF THE MARINE CYANOBACTERIUM TRICHODESMIUM IMS101 IN CONTINUOUS CULTURE: EFFECTS OF GROWTH RATE ON N2‐FIXATION RATE,BIOMASS, AND C:N:P STOICHIOMETRY1

Trichodesmium N₂ fixation has been studied for decades in situ and, recently, in controlled laboratory conditions; yet N₂‐fixation rate estimates still vary widely. This variance has made it difficult to accurately estimate the input of new nitrogen (N) by Trichodesmium to the oligotrophic gyres of the world ocean. Field and culture studies demonstrate that trace metal limitation, phosphate availability, the preferential uptake of combined N, light intensity, and temperature may all affect N₂ fixation, but the interactions between growth rate and N₂ fixation have not been well characterized in this marine diazotroph. To determine the effects of growth rate on N₂ fixation, we established phosphorus (P)–limited continuous cultures of Trichodesmium, which we maintained at nine steady‐state growth rates ranging from 0.27 to 0.67 d^?1. As growth rate increased, biomass (measured as particulate N) decreased, and N₂‐fixation rate increased linearly. The carbon to nitrogen ratio (C:N) varied from 5.5 to 6.2, with a mean of 5.8 ± 0.2 (mean ± SD, N = 9), and decreased significantly with growth rate. The N:P ratio varied from 23.4 to 45.9, with a mean of 30.5 ± 6.6 (mean ± SD, N = 9), and remained relatively constant over the range of growth rates studied. Relative constancy of C:N:P ratios suggests a tight coupling between the uptake of these three macronutrients and steady‐state growth across the range of growth rates. Our work demonstrates that growth rate must be considered when planning studies of the effects of environmental factors on N₂ fixation and when modeling the impact of Trichodesmium as a source of new N to oligotrophic regions of the ocean.     

Iron-Stimulated N2 Fixation and Growth in Natural and Cultured Populations of the Planktonic Marine Cyanobacteria Trichodesmium spp

In light of recent proposals that iron (Fe) availability may play an important role in controlling oceanic primary production and nutrient flux, its regulatory impact on N₂ fixation and production dynamics was investigated in the widespread and biogeochemically important diazotrophic, planktonic cyanobacteria Trichodesmium spp. Fe additions, as FeCl₃ and EDTA-chelated FeCl₃, enhanced N₂ fixation (nitrogenase activity), photosynthesis (CO₂ fixation), and growth (chlorophyll a production) in both naturally occurring and cultured (on unenriched oligotrophic seawater) Trichodesmium populations. Maximum enhancement of these processes occurred under FeEDTA-amended conditions. On occasions, EDTA alone led to enhancement. No evidence for previously proposed molybdenum or phosphorus limitation was found. Our findings geographically extend support for Fe limitation of N₂ fixation and primary production to tropical and subtropical oligotrophic ocean waters often characterized by Trichodesmium blooms.