Long-Term Temperature Acclimation of Photosynthesis in Steady-State Cultures of the Polar Diatom <Emphasis Type="Italic">Fragilariopsis cylindrus</Emphasis>

Cultures of the obligate psychrophilic diatom Fragilariopsis cylindrus (Grunow) were grown for 4 months under steady-state conditions at −1 °C and +7 °C (50 μmol photons m⁻² s⁻¹) prior to measurements in order to investigate long-term acclimation of photosynthesis to both temperatures. No differences in maximum intrinsic quantum yield of PS II (F_V/F_M) and relative electron transport rates could be detected at either temperature after 4 months of acclimation. Measurements of photosynthesis (relative electron transport rates) vs. irradiance (P vs. E curves) revealed similar values for relative light utilization efficiency (α = 0.57 at −1 °C, α = 0.60 at +7 °C) but higher values for irradiance levels at which photosynthesis saturates (E_K) at −1 °C and, therefore, higher maximum photosynthesis (P_MAX = 54 (relative units) at −1 °C, P_MAX = 49 at +7 °C). Nonphotochemical quenching (NPQ) measurements at 385 μmol photons m⁻² s⁻¹ indicated higher (37%) NPQ for diatoms grown at −1 °C compared to +7 °C, which was possibly related to a 2-fold increase in the concentration of the pigment diatoxanthin and a 9-fold up-regulation of a gene encoding a fucoxanthin chlorophyll a,c-binding protein. Expression of the D1 protein encoding gene psbA was ca. 1.5-fold up-regulated at −1 °C, whereas expression levels of other genes from Photosystem II (psbC, psbU, psbO), as well as rbcL, the gene encoding the Rubisco large subunit were similar at both temperatures. However, a 2-fold up-regulation of a plastid glyceraldehyde-P dehydrogenase at −1 °C indicated enhanced Calvin cycle activity. This study revealed for the first time that a polar diatom could efficiently acclimate photosynthesis over a wide range of polar temperatures given enough time. Acclimation of photosynthesis at −1 °C was probably regulated similarly to high light acclimation.     

Changes in the condition of photosynthetic apparatus of a diatom alga <Emphasis Type="Italic">Thallassiosira weisflogii</Emphasis> during photoadaptation and photodamage

Two populations of a diatom alga Thallassiosira weisflogii were grown at photon flux densities (PFD) of 0.8 and 8 μmol/(m² s). For both diatom populations, the recovery of chlorophyll fluorescence parameters (F ₀, F _m, F _v/F _m, and NPQ) was monitored after nondestructive irradiation by visible light at PFD of 40 μmol/(m² s) and after high-intensity irradiation by visible light (1000–4000 μmol/(m² s)). The exposure of diatoms to PFD of 40 μmol/(m² s)—higher than PFD used for algal growth but still nondamaging to photosynthetic apparatus—induced nonphotochemical quenching (NPQ), which was stronger in algae grown at higher PFD (8 μmol/(m² s)) than in algae grown at low light. After irradiation with high-intensity light, the recovery of chlorophyll fluorescence parameters was more pronounced in algae grown at elevated PFD level. During short-term irradiation of diatoms with high-intensity visible light (1000 μmol/(m² s)), a stronger NPQ was observed in the culture adapted to high irradiance. After the treatment of algae with dithiothreitol (an inhibitor of carotenoid deepoxidase in the diadinoxanthin cycle) or NH₄Cl (an agent abolishing the proton gradient at thylakoid membranes), a short exposure of algae to PFD of 40 μmol/(m² s) induced hardly any nonphotochemical quenching. The results indicate the dominant contribution of xanthophyll cycle carotenoids to energy-dependent quenching.     

Disentangling two non-photochemical quenching processes in Cyclotella meneghiniana by spectrally-resolved picosecond fluorescence at 77 K

Diatoms, which are primary producers in the oceans, can rapidly switch on/off efficient photoprotection to respond to fast light-intensity changes in moving waters. The corresponding thermal dissipation of excess-absorbed-light energy can be observed as non-photochemical quenching (NPQ) of chlorophyll a fluorescence. Fluorescence-induction measurements on Cyclotella meneghiniana diatoms show two NPQ processes: qE₁ relaxes rapidly in the dark while qE₂ remains present upon switching to darkness and is related to the presence of the xanthophyll-cycle pigment diatoxanthin (Dtx). We performed picosecond fluorescence measurements on cells locked in different (quenching) states, revealing the following sequence of events during full development of NPQ. At first, trimers of light-harvesting complexes (fucoxanthin–chlorophyll a/c proteins), or FCPa, become quenched, while being part of photosystem II (PSII), due to the induced pH gradient across the thylakoid membrane. This is followed by (partial) detachment of FCPa from PSII after which quenching persists. The pH gradient also causes the formation of Dtx which leads to further quenching of isolated PSII cores and some aggregated FCPa. In subsequent darkness, the pH gradient disappears but Dtx remains present and quenching partly pertains. Only in the presence of some light the system completely recovers to the unquenched state.     

Ultrafast fluorescence study on the location and mechanism of non-photochemical quenching in diatoms

The diatom algae, responsible for at least a quarter of the global photosynthetic carbon assimilation in the oceans, are capable of switching on rapid and efficient photoprotection, which helps them cope with the large fluctuations of light intensity in the moving waters. The enhanced dissipation of excess excitation energy becomes visible as non-photochemical quenching (NPQ) of chlorophyll a fluorescence. Intact cells of the diatoms Cyclotella meneghiniana and Phaeodactylum tricornutum, which show different NPQ induction kinetics under high light illumination, were investigated by picosecond time-resolved fluorescence under dark and NPQ-inducing high light conditions. The fluorescence kinetics revealed that there are two independent sites responsible for NPQ. The first quenching site is located in an FCP antenna system that is functionally detached from both photosystems, while the second quenching site is located in the PSII-attached antenna. Notwithstanding their different npq induction and reversal kinetics, both diatoms showed identical NPQ via both mechanisms in the steady-state. Their fluorescence decays in the dark-adapted states were different, however. A detailed quenching model is proposed for NPQ in diatoms.     

THE IMPACT OF NONPHOTOCHEMICAL QUENCHING OF FLUORESCENCE ON THE PHOTON BALANCE IN DIATOMS UNDER DYNAMIC LIGHT CONDITIONS1

The nonphotochemical quenching (NPQ) of fluorescence is an important photoprotective mechanism in particular under dynamic light conditions. Its photoprotective potential was suggested to be a functional trait of algal diversity. In the present study, the influence of the photoprotective capacity on the growth balance was investigated in two diatoms, which possess different NPQ characteristics. It was hypothesized that under fluctuating light conditions Cyclotella meneghiniana Kütz. would benefit from its large and flexible NPQ potential, whereas the comparably small NPQ capacity in Skeletonema costatum (Grev.) Cleve should exert an unfavorable impact on growth. The results of the study clearly falsify this hypothesis. Although C. meneghiniana possesses a fast NPQ component, this diatom was not able to recover its full NPQ capacity under fluctuating light. On the other hand, the induction of NPQ at relatively low irradiance in S. costatum resulted in rather small differences in the fraction of energy dissipation by the NPQ mechanism in the comparison of both diatoms. Larger differences were found in the metabolic characteristics. Both diatoms differed in their biomass composition, with a higher content of lipids in C. meneghiniana but higher amounts of carbohydrates in S. costatum. Finally, the lower degree of reduction in the biomass compensated for the higher respiration rates in S. costatum and resulted in a higher quantum efficiency of biomass production. An indirect correlation between the photoprotective and the metabolic capacity is discussed.     

The integrative expression of GUS gene driven by FCP promoter in the seaweed Laminaria japonica (Phaeophyta)

In this study, the background activity of β-glucuronidase (GUS) was analyzed histochemically and fluorometrically in the negative control of Laminaria japonica (Phaeophyta) thalli, showing low level of activity. GUS gene transformation without selectable gene in L. japonica was performed using four different promoters, i.e., Cauliflower mosaic virus 35S promoter (CaMV35S) from cauliflower mosaic virus, ubiquitin promoter (UBI) from maize, adenine-methyl transfer enzyme gene promoter (AMT) from virus in green alga Chlorella, and fucoxanthin chlorophyll a/c-binding protein gene promoter (FCP) from diatom Phaeodactylum tricornutum. The GUS transient activity was determined fluorometrically after bombarding sliced parthenogenetic sporophytes explants, and it was found that the activity resulting from CaMV35S and FCP promoters (in 114.3 and 80.6 pmol MU min⁻¹ (mg protein)⁻¹, respectively) was higher than for the other two promoters. The female gametophytes were bombarded and regenerated parthenogenetic sporophytes. FCP was the only promoter that resulted in detectable GUS chimeric expression activity during histochemical staining and polymerase chain reaction. Results of Southern blot showed that GUS gene was integrated with the L. japonica genome.     

Slow zeaxanthin accumulation and the enhancement of CP26 collectively contribute to an atypical non-photochemical quenching in macroalga Ulva prolifera under high light

Non-photochemical quenching (NPQ) is an important photoprotective mechanism in plants, which dissipates excess energy and further protects the photosynthetic apparatus under high light stress. NPQ can be dissected into a number of components: qE, qZ, and qI. In general, NPQ is catalyzed by two independent mechanisms, with the faster-activated quenching catalyzed by the monomeric light-harvesting complex (LHCII) proteins and the slowly activated quenching catalyzed by LHCII trimers, both processes depending on zeaxanthin but to different extent. Here, we studied the NPQ of the intertidal green macroalga, Ulva prolifera, and found that the NPQ of U. prolifera lack the faster-activated quenching, and showed much greater sensitivity to dithiothreitol (DTT) than to dicyclohexylcarbodiimide (DCCD). Further results suggested that the monomeric LHC proteins in U. prolifera included only CP29 and CP26, but lacked CP24, unlike Arabidopsis thaliana and the moss Physcomitrella patens. Moreover, the expression levels of CP26 increased significantly following exposure to high light, but the concentrations of the two important photoprotective proteins (PsbS and light-harvesting complex stress-related [LhcSR]) did not change upon the same conditions. Analysis of the xanthophyll cycle pigments showed that, upon exposure to high light, zeaxanthin synthesis in U. prolifera was gradual and much slower than that in P. patens, and could effectively be inhibited by DTT. Based on these results, we speculate the enhancement of CP26 and slow zeaxanthin accumulation provide an atypical NPQ, making this green macroalga well adapted to the intertidal environments.     

Acclimation of the diatom Stephanodiscus neoastraea and the cyanobacterium Planktothrix agardhii to simulated natural light fluctuations

Functional and structural characteristics of the photosynthetic apparatus were studied in the diatom Stephanodiscus neoastraea and the cyanobacterium Planktothrix agardhii which were grown semi-continuously under constant irradiance or under simulated natural light fluctuations. The light fluctuations consisted of 24 oscillations of exponentially increasing and decreasing irradiance over a 12-h light period. Maximum irradiance was 1100 μmol photons m⁻² s⁻¹ with the ratio of maximum to minimum intensities being 100, simulating Langmuir circulations with a ratio of euphotic to mixing depth of 1. S. neoastraea acclimated to the light fluctuations by doubling the number and halving the size of photosynthetic units (PS II) while the amount of chlorophylls and carotenoids remained unchanged. The chlorophyll-specific maximum photosynthetic rate was enhanced while the slope of the photosynthesis versus irradiance curves was not influenced by the light fluctuations. Acclimation of P. agardhii was mainly characterized by an increase in chlorophyll content. Both photosystems showed only little changes in number and size. Maximum photosynthetic rate, saturating irradiance and initial slope of the photosynthesis versus irradiance curves did not vary. Although both high and low light were contained in the fluctuating light, an analogy to low or high light acclimation was not found for the diatom nor for the cyanobacterium acclimated to light fluctuations. We suggest that the acclimation to fluctuating light is a response type outside the known scheme of low and high light acclimation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Interactive Effects of CO2, Temperature,Irradiance, and Nutrient Limitation on the Growth and Physiology of the Marine Diatom Thalassiosira pseudonana (Coscinodiscophyceae)

The marine diatom Thalassiosira pseudonana was grown in continuous culture systems to study the interactive effects of temperature, irradiance, nutrient limitation, and the partial pressure of CO₂ (pCO₂) on its growth and physiological characteristics. The cells were able to grow at all combinations of low and high irradiance (50 and 300 μmol photons · m⁻² · s⁻¹, respectively, of visible light), low and high pCO₂ (400 and 1,000 μatm, respectively), nutrient limitation (nitrate-limited and nutrient-replete conditions), and temperatures of 10–32°C. Under nutrient-replete conditions, there was no adverse effect of high pCO₂ on growth rates at temperatures of 10–25°C. The response of the cells to high pCO₂ was similar at low and high irradiance. At supraoptimal temperatures of 30°C or higher, high pCO₂ depressed growth rates at both low and high irradiance. Under nitrate-limited conditions, cells were grown at 38 ± 2.4% of their nutrient-saturated rates at the same temperature, irradiance, and pCO₂. Dark respiration rates consistently removed a higher percentage of production under nitrate-limited versus nutrient-replete conditions. The percentages of production lost to dark respiration were positively correlated with temperature under nitrate-limited conditions, but there was no analogous correlation under nutrient-replete conditions. The results suggest that warmer temperatures and associated more intense thermal stratification of ocean surface waters could lower net photosynthetic rates if the stratification leads to a reduction in the relative growth rates of marine phytoplankton, and at truly supraoptimal temperatures there would likely be a synergistic interaction between the stresses from temperature and high pCO₂ (lower pH).     

The importance of a highly active and DeltapH-regulated diatoxanthin epoxidase for the regulation of the PS II antenna function in diadinoxanthin cycle containing algae

The present study focuses on the regulation of diatoxanthin (Dtx) epoxidation in the diadinoxanthin (Ddx) cycle containing algae Phaeodactylum tricornutum, Thalassiosira pseudonana, Cyclotella meneghiniana and Prymnesium parvum and its significance for the control of the photosystem II (PS II) antenna function. Our data show that Dtx epoxidase can exhibit extremely high activities when algal cells are transferred from high light (HL) to low light (LL). Under HL conditions, Dtx epoxidation is strongly inhibited by the light-driven proton gradient. Uncoupling of the cells during HL illumination restores the high epoxidation rates observed during LL. In Ddx cycle containing algae, non-photochemical quenching of chlorophyll fluorescence (NPQ) is directly correlated with the Dtx concentration and independent of the presence of the proton gradient. This means that a fast conversion of PS II from the heat dissipating state back to the light-harvesting state can only be realized by an efficient removal of the quenching pigment Dtx. It is proposed that the high Dtx epoxidation rates during LL illumination serve exactly this purpose. The inhibition of Dtx epoxidation by the DeltapH, on the other hand, ensures rapid increases in the Dtx concentration when photoprotection under conditions of HL illumination is required. The regulation of the PS II antenna function in Ddx cycle containing algae is different to that in violaxanthin (Vx) cycle containing plants, where for the zeaxanthin (Zx)-dependent NPQ the presence of a proton gradient is mandatory. In the green alga Chlorella vulgaris conversion of PS II from the heat dissipating state back to the light-harvesting state is controlled by the DeltapH, whose relaxation after a transition from HL to darkness or LL rapidly abolishes the thermal dissipation of excitation energy, including the Zx-dependent NPQ. Due to the inability of Zx to quench fluorescence in the absence of the DeltapH a fast epoxidation of Zx to Vx in LL is not needed and is missing in Chlorella vulgaris.     

Depth distribution of photosynthetic pigments and diatoms in the sediments of a microtidal fjord

The depth distribution of photosynthetic pigments and benthic marine diatoms was investigated in late spring at three different sites on the Swedish west coast. At each site, sediment cores were taken at six depths (7–35 m) by scuba divers. It was hypothesized that (1) living benthic diatoms constitute a substantial part of the benthic microflora even at depths where the light levels are <1% of the surface irradiance, and (2) the changing light environment along the depth gradient will be reflected in (a) the composition of diatom assemblages, and (b) different pigment ratios. Sediment microalgal communities were analysed using epifluorescence microscopy (to study live cells), light microscopy and scanning electron microscopy (diatom preparations), and HPLC (photosynthetic pigments). Pigments were calculated as concentrations (mg m^–2) and as ratios relative to chlorophyll a. Hypothesis (1) was accepted. At 20 m, the irradiance was 0.2% of surface irradiance and at 7 m, 1%. Living (epifluorescent) benthic diatoms were found down to 20 m at all sites. The cell counts corroborated the diatom pigment concentrations, decreasing with depth from 7 to 25 m, levelling out between 25 and 35 m. There were significant positive correlations between chlorophyll a and living (epifluorescent) benthic diatoms and between the diatom pigment fucoxanthin and chlorophyll a. Hypothesis (2) was only partly accepted because it could not be shown that light was the main environmental factor. A principal component analysis on diatom species showed that pelagic forms characterized the deeper locations (25–35 m), and epipelic–epipsammic taxa the shallower sites (7–20 m). Redundancy analyses showed a significant relationship between diatom taxa and environmental factors – temperature, salinity, and light intensities explained 57% of diatom taxa variations.     

Photosynthetic performance and light response of two olive cultivars under different water and light regimes

The olive tree (Olea europaea L.) is commonly grown in the Mediterranean area, where it is adapted to resist periods characterized by severe drought and high irradiance levels. Photosynthetic efficiency (in terms of F_v/F_m and Φ_PSII), photochemical (q_P) and nonphotochemical quenching (NPQ) were determined in two-year-old olive plants (cultivars Coratina and Biancolilla) grown under two different light levels (exposed plants, EP, and shaded plants, SP) during a 21-day controlled water deficit. After reaching the maximum level of drought stress, plants were rewatered for 23 days. During the experimental period, measurements of gas exchange and chlorophyll (Chl) fluorescence were carried out to study the photosynthetic performance of olive plants. The synergical effect of drought stress and high irradiance levels caused a reduction of gas exchange and photosynthetic efficiency and these decreases were more marked in EP. EP showed a higher degree of photoinhibition, a higher NPQ and a lower q_P if compared to SP. Coratina was more sensitive to high light and drought stress but also showed a slower recovery during rewatering, whereas Biancolilla showed a less marked photosynthesis depression during drought and a considerable resilience during rewatering. The results confirm that photoinhibition due to high light intensity and water deficit can be an important factor that affects photosynthetic productivity in this species.     

High salt stress in the upper part of floating mats of Ulva prolifera,a species that causes green tides,enhances non‐photochemical quenching

Salt stress is a major abiotic stress factor that can induce many adverse effects on photosynthetic organisms. Plants and algae have developed several mechanisms that help them respond to adverse environments. Non‐photochemical quenching (NPQ) is one of these mechanisms. The thalli of algae in the intertidal zone that are attached to rocks can be subjected to salt stress for a short period of time due to the rise and fall of the tide. Ulva prolifera causes green tides and can form floating mats when green tides occur and the upper part of the thalli is subjected to high salt stress for a long period of time. In this study, we compared the Ulva prolifera photosynthetic activities and NPQ kinetics when it is subjected to different salinities over various periods of time. Thalli exposed to a salinity of 90 for 4 d showed enhanced NPQ, and photosynthetic activities decreased from 60 min after exposure up to 4 d. This indicated that the induction of NPQ in Ulva prolifera under salt stress was closely related to the stressing extent and stressing time. The enhanced NPQ in the treated samples exposed for 4 d may explain why the upper layer of the floating mats formed by Ulva prolifera thalli were able to survive in the harsh environment. Further inhibitor experiments demonstrated that the enhanced NPQ was xanthophyll cycle and transthylakoid proton gradient‐dependent. However, photosystem II subunit S and light‐harvesting complex stress‐related protein didn't over accumulate and may not be responsible for the enhanced NPQ.     

Photoadaptation of two members of the Chlorophyta (Scenedesmus and Chlorella) in laboratory and outdoor cultures: changes in chlorophyll fluorescence quenching and the xanthophyll cycle

The role of the xanthophyll cycle in the adaptation of two chlorococcal algae Scenedesmus quadricauda and Chlorella sorokiniana to high irradiance was studied under laboratory and outdoor conditions. We wished to elucidate whether the xanthophyll cycle plays a key role in dissipating the excesses of absorbed light, as in higher plants, and to characterise the relationship between chlorophyll fluorescence parameters and the content of xanthophyll-cycle pigments. The xanthophyll cycle was found to be operative in both species; however, its contribution to overall non-photochemical quenching (NPQ) could only be distinguished in Scenedesmus (15–20% of total NPQ). The Scenedesmus cultures showed a larger pool of xanthophyll-cycle pigments than Chlorella, and lower sensitivity to photoinhibition as judged from the reduction of maximum quantum yield of photosystem II. In general, both algae had a larger xanthophyll-cycle pool when grown outdoors than in laboratory cultures. Comparing the two species, Scenedesmus exhibited a higher capacity to adapt to high irradiance, due to an effective quenching mechanism and high photosynthetic capacity; in contrast, Chlorella represents a species with a larger antennae system, less-efficient quenching and lower photosynthetic performance. Non-photochemical quenching (NPQ) induced through the xanthophyll cycle can, to a limited extent, represent a regulatory factor in diluted algal cultures grown in outdoor solar photobioreactors, as well as in natural algal phytoplankton populations exposed transiently to high irradiance. However, it does not play an appreciable role in dense, well-mixed microalgal suspensions. Received: 6 August 1998 / Accepted: 12 February 1999     

Photoprotection in a zeaxanthin- and lutein-deficient double mutant of Arabidopsis

When light absorption by a plant exceeds its capacity for light utilization, photosynthetic light harvesting is rapidly downregulated by photoprotective thermal dissipation, which is measured as nonphotochemical quenching of chlorophyll fluorescence (NPQ). To address the involvement of specific xanthophyll pigments in NPQ, we have analyzed mutants affecting xanthophyll metabolism in Arabidopsis thaliana. An npq1 lut2 double mutant was constructed, which lacks both zeaxanthin and lutein due to defects in the violaxanthin de-epoxidase and lycopene -cyclase genes. The npq1 lut2 strain had normal Photosystem II efficiency and nearly wild-type concentrations of functional Photosystem II reaction centers, but the rapidly reversible component of NPQ was completely inhibited. Despite the defects in xanthophyll composition and NPQ, the npq1 lut2 mutant exhibited a remarkable ability to tolerate high light.This revised version was published online in October 2005 with corrections to the Cover Date.     

Effect of temperature on light-limited growth of the harmful diatom Eucampia zodiacus Ehrenberg, a causative organism in the discoloration of Porphyra thalli

The diatom Eucampia zodiacus Ehrenberg is one of the harmful diatoms which indirectly cause, through nutrient depletion, discoloration of Porphyra thalli. The effect of temperature on light-limited growth of E. zodiacus was examined at 13 irradiance levels (5–350 μmol m⁻² s⁻¹) in combination with five temperatures (8.0–25.0 °C). The results showed that all the parameters of growth-irradiance curves, such as the maximum growth rate (μ_m), half saturation constant (K_s), threshold value of irradiance (I₀) and saturation irradiance for growth (S), increased with increasing temperature. On the basis of the relationship between temperature and growth-irradiance curves and seasonal fluctuation of the light environment in Harima-Nada, the effect of irradiance on the population dynamics of E. zodiacus during the period from October to March was evaluated using two indices, depth of the threshold irradiance for growth (D_t) and depth where a half of its maximum growth rate is attained (D_k). D_t and D_k remained almost stable from October to December, but gradually increased in early March. This indicates that the range of depth at which E. zodiacus was able to grow increased markedly in early spring when E. zodiacus blooms in Harima-Nada. As the vegetative cells of E. zodiacus tend to distribute in relatively deeper water layers, where growth is limited by irradiance, the increase in the depth range over which E. zodiacus is able to grow is concluded to be an important factor allowing development of its blooms.