Inactivation Of Photosystem II Research Articles

Effect of Mycotoxin Cytochalasin A on Photosystem II in Ageratina adenophora.

Biological herbicides have received much attention due to their abundant resources, low development cost, unique targets and environmental friendliness. This study reveals some interesting effects of mycotoxin cytochalasin A (CA) on photosystem II (PSII). Our results suggested that CA causes leaf lesions on Ageratina adenophora due to its multiple effects on PSII. At a half-inhibitory concentration of 58.5 μΜ (I50, 58.5 μΜ), the rate of O2 evolution of PSII was significantly inhibited by CA. This indicates that CA possesses excellent phytotoxicity and exhibits potential herbicidal activity. Based on the increase in the J-step of the chlorophyll fluorescence rise OJIP curve and the analysis of some JIP-test parameters, similar to the classical herbicide diuron, CA interrupted PSII electron transfer beyond QA at the acceptor side, leading to damage to the PSII antenna structure and inactivation of reaction centers. Molecular docking model of CA and D1 protein of A. adenophora further suggests that CA directly targets the QB site of D1 protein. The potential hydrogen bonds are formed between CA and residues D1-His215, D1-Ala263 and D1-Ser264, respectively. The binding of CA to residue D1-Ala263 is novel. Thus, CA is a new natural PSII inhibitor. These results clarify the mode of action of CA in photosynthesis, providing valuable information and potential implications for the design of novel bioherbicides.

Dissecting photosynthetic electron transport and photosystems performance in Jerusalem artichoke (Helianthus tuberosus L.) under salt stress.

Jerusalem artichoke (Helianthus tuberosus L.), a vegetable with medical applications, has a strong adaptability to marginal barren land, but the suitability as planting material in saline land remains to be evaluated. This study was envisaged to examine salt tolerance in Jerusalem artichoke from the angle of photosynthetic apparatus stability by dissecting the photosynthetic electron transport process. Potted plants were exposed to salt stress by watering with a nutrient solution supplemented with NaCl. Photosystem I (PSI) and photosystem II (PSII) photoinhibition appeared under salt stress, according to the significant decrease in the maximal photochemical efficiency of PSI (△MR/MR0) and PSII. Consistently, leaf hydrogen peroxide (H2O2) concentration and lipid peroxidation were remarkably elevated after 8 days of salt stress, confirming salt-induced oxidative stress. Besides photoinhibition of the PSII reaction center, the PSII donor side was also impaired under salt stress, as a K step emerged in the prompt chlorophyll transient, but the PSII acceptor side was more vulnerable, considering the decreased probability of an electron movement beyond the primary quinone (ETo/TRo) upon depressed upstream electron donation. The declined performance of entire PSII components inhibited electron inflow to PSI, but severe PSI photoinhibition was not averted. Notably, PSI photoinhibition elevated the excitation pressure of PSII (1-qP) by inhibiting the PSII acceptor side due to the negative and positive correlation of △MR/MR0 with 1-qP and ETo/TRo, respectively. Furthermore, excessive reduction of PSII acceptors side due to PSI photoinhibition was simulated by applying a specific inhibitor blocking electron transport beyond primary quinone, demonstrating that PSII photoinhibition was actually accelerated by PSI photoinhibition under salt stress. In conclusion, PSII and PSI vulnerabilities were proven in Jerusalem artichoke under salt stress, and PSII inactivation, which was a passive consequence of PSI photoinhibition, hardly helped protect PSI. As a salt-sensitive species, Jerusalem artichoke was recommended to be planted in non-saline marginal land or mild saline land with soil desalination measures.

Drought stress modulates expression of aquaporin gene and photosynthetic efficiency in Pennisetum glaucum (L.) R. Br. genotypes

Photosynthetic efficiency and availability of water have a close relationship which determines the growth and development of plants. Drought has a remarkable effect on photosynthetic efficiency by affecting the functioning of Photosystem II (PSII). In this study, two drought genotypes of pearl millet were taken and assessed for photosynthetic performance of PSII (electron transport rate (ETR) and maximum yield) under a simulated drought stress condition. Drought stress caused a gradual downregulation in the chlorophyll content in both genotypes. The elevated content of β-carotene was recorded on the 12th day of the experiment. The photo inactivation of PSII was also assessed in both the genotypes. On the basis of an overall comparison, it is evident that the genotype IP14599 has better adapted to drought than the genotype IP14222. Furthermore, the results were confirmed by the analysis of transcript expression level of two aquaporin candidate genes (PIP2;3 and PIP1;5) and it was found that genotype IP14599 was better to maintain homeostasis by increasing in water channeling and movement in response to drought as compared to IP14222. Therefore, it was confirmed that PIP2; 3 and PIP1;5 gene modulate expression of mRNA to provide tolerance against drought in millet plants.

Seasonal Changes in Primary Photosynthetic Events during Low Temperature Adaptation of Pinus sylvestris in Central Yakutia

The methods of chlorophyll fluorescence induction and HPLC were used to study the influence of autumnal temperature decrease on photochemical electron-transport activity of photosystem II (PSII), nonphotochemical quenching of excessive excitation energy, and the composition of pigments in the firstyear needles of Pinus sуlvestris L. trees grown naturally in Central Yakutia. In the period from the beginning of September to October 10, the chlorophyll content was reduced by half, while the Chl a/b ratio increased from 2.9 to 4.3–4.5, indicating the degradation of peripheral antenna complexes. The decrease in average daily temperature to 4.9–6.4°C led to a transient increase in the quantum yield of nonphotochemical quenching (ΔpH-dependent parameter φNPQ). These changes were accompanied by a slow accumulation of unregulated zeaxanthin fraction insensitive to illumination conditions. The further decrease in average daily temperature to near-zero levels was paralleled by a sharp increase in zeaxanthin content, while the pH-dependent quenching was replaced with the constitutive quenching (parameter φf,D) because of the supposed structural reorganization of PSII. These processes were accompanied by a fast decrease in PSII functional activity, which was mostly due to the impairment of plastoquinone photochemical reduction. Freezing temperatures (from–3.6 to–12.1) destructed the oxygen-evolving complex in PSII and completely inactivated the PSII reaction centers. It is concluded that the largest changes in the condition of photosynthetic apparatus occur at a near-zero temperature range and proceed until complete inactivation of PSII under the action of freezing temperatures.

Light acclimation of shade-tolerant and sun-resistant Tradescantia species: photochemical activity of PSII and its sensitivity to heat treatment.

In this work, we have compared photosynthetic characteristics of photosystem II (PSII) in Tradescantia leaves of two contrasting ecotypes grown under the low light (LL) and high light (HL) regimes during their entire growth period. Plants of the same genus, T. fluminensis (shade-tolerant) and T. sillamontana (sun-resistant), were cultivated at 50-125µmol photons m-2s-1 (LL) or at 875-1000µmol photons m-2s-1 (HL). Analyses of intrinsic PSII efficiency was based on measurements of fast chlorophyll (Chl) a fluorescence kinetics (the OJIP test). The fluorescence parameters Fv/Fm (variable fluorescence) and F0 (the initial level of fluorescence) in dark-adapted leaves were used to quantify the photochemical properties of PSII. Plants of different ecotypes showed different sustainability with respect to changes in the environmental light intensity and temperature treatment. The sun-resistant species T. sillamontana revealed the tolerance to variations in irradiation intensity, demonstrating constancy of maximum quantum efficiency of PSII upon variations of the growth light. In contrast to T. sillamontana, facultative shade species T. fluminensis demonstrated variability of PSII photochemical activity, depending on the growth light intensity. The susceptibility of T. fluminensis to solar stress was documented by a decrease in Fv/Fm and a rise of F0 during the long-term exposition of T. fluminensis to HL, indicating the loss of photochemical activity of PSII. The short-term (10min) heat treatment of leaf cuttings caused inactivation of PSII. The temperature-dependent heating effects were different in T. fluminensis and T. sillamontana. Sun-resistant plants T. sillamontana acclimated to LL and HL displayed the same plots of Fv/Fm versus the treatment temperature (t), demonstrating a decrease in Fv/Fm at t ≥ 45°C. The leaves of shadow-tolerant species T. fluminensis grown under the LL and HL conditions revealed different sensitivities to heat treatment. Plants grown under the solar stress conditions (HL) demonstrated a gradual decline of Fv/Fm at lower heating temperatures (t ≥ 25°C), indicating the "fragility" of their PSII as compared to T. fluminensis grown at LL. Different responses of sun and shadow species of Tradescantia to growth light and heat treatment are discussed in the context of their biochemical and ecophysiological properties.

The mechanism of photosystem-II inactivation during sulphur deprivation-induced H2 production in Chlamydomonas reinhardtii.

Sulphur limitation may restrain cell growth and viability. In the green alga Chlamydomonas reinhardtii, sulphur limitation may induce H2 production lasting for several days, which can be exploited as a renewable energy source. Sulphur limitation causes a large number of physiological changes, including the inactivation of photosystemII (PSII), leading to the establishment of hypoxia, essential for the increase in hydrogenase expression and activity. The inactivation of PSII has long been assumed to be caused by the sulphur-limited turnover of its reaction center protein PsbA. Here we reinvestigated this issue in detail and show that: (i) upon transferring Chlamydomonas cells to sulphur-free media, the cellular sulphur content decreases only by about 25%; (ii) as demonstrated by lincomycin treatments, PsbA has a significant turnover, and other photosynthetic subunits, namely RbcL and CP43, are degraded more rapidly than PsbA. On the other hand, sulphur limitation imposes oxidative stress early on, most probably involving the formation of singlet oxygen in PSII, which leads to an increase in the expression of GDP-L-galactose phosphorylase, playing an essential role in ascorbate biosynthesis. When accumulated to the millimolar concentration range, ascorbate may inactivate the oxygen-evolving complex and provide electrons to PSII, albeit at a low rate. In the absence of a functional donor side and sufficient electron transport, PSII reaction centers are inactivated and degraded. We therefore demonstrate that the inactivation of PSII is a complex and multistep process, which may serve to mitigate the damaging effects of sulphur limitation.

Photosystem II activity of wild type Synechocystis PCC 6803 and its mutants with different plastoquinone pool redox states

To assess the role of redox state of photosystem II (PSII) acceptor side electron carriers in PSII photochemical activity, we studied sub-millisecond fluorescence kinetics of the wild type Synechocystis PCC 6803 and its mutants with natural variability in the redox state of the plastoquinone (PQ) pool. In cyanobacteria, dark adaptation tends to reduce PQ pool and induce a shift of the cyanobacterial photosynthetic apparatus to State 2, whereas illumination oxidizes PQ pool, leading to State 1 (Mullineaux, C. W., and Holzwarth, A. R. (1990) FEBS Lett., 260, 245-248). We show here that dark-adapted Ox(-) mutant with naturally reduced PQ is characterized by slower QA(-) reoxidation and O2 evolution rates, as well as lower quantum yield of PSII primary photochemical reactions (Fv/Fm) as compared to the wild type and SDH(-) mutant, in which the PQ pool remains oxidized in the dark. These results indicate a large portion of photochemically inactive PSII reaction centers in the Ox(-) mutant after dark adaptation. While light adaptation increases Fv/Fm in all tested strains, indicating PSII activation, by far the greatest increase in Fv/Fm and O2 evolution rates is observed in the Ox(-) mutant. Continuous illumination of Ox(-) mutant cells with low-intensity blue light, that accelerates QA(-) reoxidation, also increases Fv/Fm and PSII functional absorption cross-section (590 nm); this effect is almost absent in the wild type and SDH(-) mutant. We believe that these changes are caused by the reorganization of the photosynthetic apparatus during transition from State 2 to State 1. We propose that two processes affect the PSII activity during changes of light conditions: 1) reversible inactivation of PSII, which is associated with the reduction of electron carriers on the PSII acceptor side in the dark, and 2) PSII activation under low light related to the increase in functional absorption cross-section at 590 nm.

Conversion of photosystem II dimer to monomers during photoinhibition is tightly coupled with decrease in oxygen-evolving activity in the diatom Chaetoceros gracilis.

The rapid turnover of photosystem II (PSII) in diatoms is thought to be at an exceptionally high rate compared with other oxyphototrophs; however, its molecular mechanisms are largely unknown. In this study, we examined the photodamage and repair processes of PSII in the marine centric diatom Chaetoceros gracilis incubated at 30 or 300μmol photons m-2s-1 in the presence of a de novo protein-synthesis inhibitor. When de novo protein synthesis was blocked by chloramphenicol (Cm), oxygen-evolving activity gradually decreased even at 30μmol photons m-2s-1 and could not be detected at 12h. PSII inactivation was enhanced by higher illumination. Using Cm-treated cells, the conversion of PSII dimer to monomers was observed by blue native PAGE. The rate of PSII monomerization was very similar to that of the decrease in oxygen-evolving activity under both light conditions. Immunological detection of D1 protein in the Cm-treated cells showed that the rate of D1 degradation was slower than that of the former two events, although it was more rapid than that observed in other oxyphototrophs. Thus, the three accelerated events, especially PSII monomerization, appear to cause the unusually high rate of PSII turnover in diatoms.

The Arabidopsis thylakoid transporter PHT4;1 influences phosphate availability for ATP synthesis and plant growth.

The Arabidopsis phosphate transporter PHT4;1 was previously localized to the chloroplast thylakoid membrane. Here we investigated the physiological consequences of the absence of PHT4;1 for photosynthesis and plant growth. In standard growth conditions, two independent Arabidopsis knockout mutant lines displayed significantly reduced leaf size and biomass but normal phosphorus content. When mutants were grown in high-phosphate conditions, the leaf phosphorus levels increased and the growth phenotype was suppressed. Photosynthetic measurements indicated that in the absence of PHT4;1 stromal phosphate was reduced to levels that limited ATP synthase activity. This resulted in reduced CO2 fixation and accumulation of soluble sugars, limiting plant growth. The mutants also displayed faster induction of non-photochemical quenching than the wild type, in line with the increased contribution of ΔpH to the proton-motive force across thylakoids. Small-angle neutron scattering showed a smaller lamellar repeat distance, whereas circular dichroism spectroscopy indicated a perturbed long-range order of photosystem II (PSII) complexes in the mutant thylakoids. The absence of PHT4;1 did not alter the PSII repair cycle, as indicated by wild-type levels of phosphorylation of PSII proteins, inactivation and D1 protein degradation. Interestingly, the expression of genes for several thylakoid proteins was downregulated in the mutants, but the relative levels of the corresponding proteins were either not affected or could not be discerned. Based on these data, we propose that PHT4;1 plays an important role in chloroplast phosphate compartmentation and ATP synthesis, which affect plant growth. It also maintains the ionic environment of thylakoids, which affects the macro-organization of complexes and induction of photoprotective mechanisms.

Lethal and sub-lethal chronic effects of the herbicide diuron on seagrass

Photosystem II herbicides from agricultural sources have been detected throughout nearshore tropical habitats including seagrass meadows. While PSII herbicides have been shown to inhibit growth in microalgae at low concentrations, the potential impacts of chronic low concentration exposures to seagrass health and growth have not been investigated. Here we exposed two tropical seagrass species Halodule uninervis and Zostera muelleri to elevated diuron concentrations (from 0.3 to 7.2μgl−1) over a 79-day period followed by a 2-week recovery period in uncontaminated seawater. PAM fluorometry demonstrated rapid effect of diuron on photosystem II (PSII) in both seagrass species at 0.3μgl−1. This effect included significant inhibition of photosynthetic efficiency (ΔF/Fm′) and inactivation of PSII (Fv/Fm) over the 11 week exposure period. Significant mortality and reductions in growth was only observed at the highest exposure concentration of 7.2μgl−1 diuron. However, biochemical indicators demonstrated that the health of seagrass after this prolonged exposure was significantly compromised at lower concentrations. For example, the drop in C:N ratios (0.6μgl−1) and reduced δ13C (1.7μgl−1) in seagrass leaves indicated reduced C-assimilation from photosynthesis. Critically, the energetic reserves of the plants (as measured by starch content in the root-rhizome complex) were approximately halved following diuron exposure at and above 1.7μgl−1. During the 2-week recovery period, the photosynthetic capacity of the seagrass improved with only plants from the highest diuron treatment still exhibiting chronic damage to PSII. This study shows that, although seagrass may survive prolonged herbicide exposures, concentrations ≥0.6μgl−1 diuron equivalents cause measureable impacts on energetic status that may leave the plants vulnerable to other simultaneous stressors. For example, tropical seagrasses have been heavily impacted by reduced light from coastal flood plumes and the effects on plant energetics from light limitation and diuron exposure (highest in flood plumes) are very similar, potentially leading to cumulative negative effects.

Chloroplast thylakoid structure in evergreen leaves employing strong thermal energy dissipation

In nature, photosynthetic organisms cope with highly variable light environments – intensities varying over orders of magnitudes as well as rapid fluctuations over seconds-to-minutes – by alternating between (a) highly effective absorption and photochemical conversion of light levels limiting to photosynthesis and (b) powerful photoprotective thermal dissipation of potentially damaging light levels exceeding those that can be utilized in photosynthesis. Adjustments of the photosynthetic apparatus to changes in light environment involve biophysical, biochemical, and structural adjustments. We used electron micrographs to assess overall thylakoid grana structure in evergreen species that exhibit much stronger maximal levels of thermal energy dissipation than the more commonly studied annual species. Our findings indicate an association between partial or complete unstacking of thylakoid grana structure and strong reversible thermal energy dissipation that, in contrast to what has been reported for annual species with much lower maximal levels of energy dissipation, is similar to what is seen under photoinhibitory conditions. For a tropical evergreen with tall grana stacks, a loosening, or vertical unstacking, of grana was seen in sun-grown plants exhibiting pronounced pH-dependent, rapidly reversible thermal energy dissipation as well as for sudden low-to-high-light transfer of shade-grown plants that responded with photoinhibition, characterized by strong dark-sustained, pH-independent thermal energy dissipation and photosystem II (PSII) inactivation. On the other hand, full-sun exposed subalpine confers with rather short grana stacks transitioned from autumn to winter via conversion of most thylakoids from granal to stromal lamellae concomitant with photoinhibitory photosynthetic inactivation and sustained thermal energy dissipation. We propose that these two types of changes (partial or complete unstacking of grana) in thylakoid arrangement are both associated with the strong non-photochemical quenching (NPQ) of chlorophyll fluorescence (a measure of photoprotective thermal energy dissipation) unique to evergreen species rather than with PSII inactivation per se.

Photoprotective mechanisms in photosystem II of Ephedra monosperma during development of frost tolerance

Dissipation of light energy absorbed by photosystem II (PSII) in assimilating shoots of an ever� green shrub Ephedra monosperma was investigated during its transition from the vegetative to frosttolerant state under natural conditions of Central Yakutia. The dynamics of modulated chlorophyll fluorescence and carotenoid content was analyzed during seasonal decrease in ambient temperature. The seasonal cooling was accompanied by a stepwise decrease in photochemical activity of PSII (Fv/Fm = (Fm - F0)/Fm). The decrease in Fv/Fm occurred from the beginning of September to the end of October, when the temperature was lowered from 10 to -8°C. During winter period the residual activity of PSII was retained at about 30% of the summer values. The seasonal decrease in temperature was accompanied by a significant stimulation of pHindepen� dent dissipative processes in reaction centers and antenna of PSII. The increase in energy losses was paral� leled by a proportional increase in zeaxanthin content on the background of decreasing content of violaxan� thin and β�carotene as possible zeaxanthin precursors. At the same time, inhibition of lightinduced non� photochemical quenching in the PSII antenna was observed. The results suggest that principal photoprotec� tive mechanisms during seasonal lowering of temperature are: (1) inactivation of PSII and dissipation of exci� tation energy in PSII reaction centers and (2) zeaxanthinmediated ener gy dissipation in the antenna com� plexes. The first mechanism seems to prevail at early stages of seasonal cooling, whereas both mechanisms are recruited from the onset of sustained freezing temperatures.

Epidermal coumaroyl anthocyanins protect sweet basil against excess light stress: multiple consequences of light attenuation

The putative photoprotective role of foliar anthocyanins continues to attract heated debate. Strikingly different experimental set-ups coupled with a poor knowledge of anthocyanin identity have likely contributed to such disparate opinions. Here, the photosynthetic responses to 30 or 100% solar irradiance were compared in two cultivars of basil, the green-leafed Tigullio (TG) and the purple-leafed Red Rubin (RR). Coumaroyl anthocyanins in RR leaf epidermis significantly mitigated the effects of high light stress. In full sunlight, RR leaves displayed several shade-plant traits; they transferred less energy than did TG to photosystem II (PSII), and non-photochemical quenching was lower. The higher xanthophyll cycle activity in TG was insufficient to prevent inactivation of PSII in full sunlight. However, TG was the more efficient in the shade; RR was far less able to accommodate a large change in irradiance. Investment of carbon to phenylpropanoid biosynthesis was more in RR than in TG in the shade, and was either greatly enhanced in TG or varied little in RR because of high sunlight. The metabolic cost of photoprotection was lower whereas light-induced increase in biomass production was higher in RR than in TG, thus making purple basil the more light tolerant. Purple basil appears indeed to display the conservative resource-use strategy usually observed in highly stress tolerant species. We conclude that the presence of epidermal coumaroyl anthocyanins confers protective benefits under high light, but it is associated with a reduced plasticity to accommodate changing light fluxes as compared with green leaves.

Thermal Acclimation of the Symbiotic Alga Symbiodinium spp. Alleviates Photobleaching under Heat Stress

A moderate increase in seawater temperature causes coral bleaching, at least partially through photobleaching of the symbiotic algae Symbiodinium spp. Photobleaching of Symbiodinium spp. is primarily associated with the loss of light-harvesting proteins of photosystem II (PSII) and follows the inactivation of PSII under heat stress. Here, we examined the effect of increased growth temperature on the change in sensitivity of Symbiodinium spp. PSII inactivation and photobleaching under heat stress. When Symbiodinium spp. cells were grown at 25°C and 30°C, the thermal tolerance of PSII, measured by the thermal stability of the maximum quantum yield of PSII in darkness, was commonly enhanced in all six Symbiodinium spp. tested. In Symbiodinium sp. CCMP827, it took 6 h to acquire the maximum PSII thermal tolerance after transfer from 25°C to 30°C. The effect of increased growth temperature on the thermal tolerance of PSII was completely abolished by chloramphenicol, indicating that the acclimation mechanism of PSII is associated with the de novo synthesis of proteins. When CCMP827 cells were exposed to light at temperature ranging from 25°C to 35°C, the sensitivity of cells to both high temperature-induced photoinhibition and photobleaching was ameliorated by increased growth temperatures. These results demonstrate that thermal acclimation of Symbiodinium spp. helps to improve the thermal tolerance of PSII, resulting in reduced inactivation of PSII and algal photobleaching. These results suggest that whole-organism coral bleaching associated with algal photobleaching can be at least partially suppressed by the thermal acclimation of Symbiodinium spp. at higher growth temperatures.

Photosystem II Function and Dynamics in Three Widely Used Arabidopsis thaliana Accessions

Columbia-0 (Col-0), Wassilewskija-4 (Ws-4), and Landsberg erecta-0 (Ler-0) are used as background lines for many public Arabidopsis mutant collections, and for investigation in laboratory conditions of plant processes, including photosynthesis and response to high-intensity light (HL). The photosystem II (PSII) complex is sensitive to HL and requires repair to sustain its function. PSII repair is a multistep process controlled by numerous factors, including protein phosphorylation and thylakoid membrane stacking. Here we have characterized the function and dynamics of PSII complex under growth-light and HL conditions. Ws-4 displayed 30% more thylakoid lipids per chlorophyll and 40% less chlorophyll per carotenoid than Col-0 and Ler-0. There were no large differences in thylakoid stacking, photoprotection and relative levels of photosynthetic complexes among the three accessions. An increased efficiency of PSII closure was found in Ws-4 following illumination with saturation flashes or continuous light. Phosphorylation of the PSII D1/D2 proteins was reduced by 50% in Ws-4 as compared to Col-0 and Ler-0. An increase in abundance of the responsible STN8 kinase in response to HL treatment was found in all three accessions, but Ws-4 displayed 50% lower levels than Col-0 and Ler-0. Despite this, the HL treatment caused in Ws-4 the lagest extent of PSII inactivation, disassembly, D1 protein degradation, and the largest decrease in the size of stacked thylakoids. The dilution of chlorophyll-protein complexes with additional lipids and carotenoids in Ws-4 may represent a mechanism to facilitate lateral protein traffic in the membrane, thus compensating for the lack of a full complement of STN8 kinase. Nevertheless, additional PSII damage occurs in Ws-4, which exceeds the D1 protein synthesis capacity, thus leading to enhanced photoinhibition. Our findings are valuable for selection of appropriate background line for PSII characterization in Arabidopsis mutants, and also provide the first insights into natural variation of PSII protein phosphorylation.

Role of Novel Dimeric Photosystem II (PSII)-Psb27 Protein Complex in PSII Repair

The multisubunit membrane protein complex Photosystem II (PSII) catalyzes one of the key reactions in photosynthesis: the light-driven oxidation of water. Here, we focus on the role of the Psb27 assembly factor, which is involved in biogenesis and repair after light-induced damage of the complex. We show that Psb27 is essential for the survival of cyanobacterial cells grown under stress conditions. The combination of cold stress (30 °C) and high light stress (1000 μmol of photons × m(-2) × s(-1)) led to complete inhibition of growth in a Δpsb27 mutant strain of the thermophilic cyanobacterium Thermosynechococcus elongatus, whereas wild-type cells continued to grow. Moreover, Psb27-containing PSII complexes became the predominant PSII species in preparations from wild-type cells grown under cold stress. Two different PSII-Psb27 complexes were isolated and characterized in this study. The first complex represents the known monomeric PSII-Psb27 species, which is involved in the assembly of PSII. Additionally, a novel dimeric PSII-Psb27 complex could be allocated in the repair cycle, i.e. in processes after inactivation of PSII, by (15)N pulse-label experiments followed by mass spectrometry analysis. Comparison with the corresponding PSII species from Δpsb27 mutant cells showed that Psb27 prevented the release of manganese from the previously inactivated complex. These results indicate a more complex role of the Psb27 protein within the life cycle of PSII, especially under stress conditions.

Photosystem II breakdown induced by reactive oxygen species in freshly-isolated Symbiodinium from Montipora (Scleractinia; Acroporidae)

Freshly-isolated Symbiodinium (FIS) have been used to study cnidarian–alga symbiosis based on the assumption that their physiological performance is comparable to that of the algae in hospite. This assumption was tested with 15 species of scleractinian corals, using pulse-amplitude modulation (PAM) chlorophyll a (chl a) fluorescence to compare maximum quantum yields (Fv/Fm) of photosystem II (PSII) in Symbiodinium in hospite and after isolation in seawater. FIS from Montipora spp. exhibited rapid and dramatic decreases (by up to >95%) in PSII activity within 30 min of isolation. In contrast, PSII activities of FIS from 8 other coral species decreased by only 5 to 21% after >4 h in seawater. To investigate possible reasons for this variation, the variation in Symbiodinium genetic type (ITS-2 types), transmission modes and several physiological indices were considered. The rapid loss of PSII activity in FIS from Montipora spp. was not correlated with Symbiodinium ITS-2 type, the mode of symbiont transmission, or ionic regulation capability, nor could consistent chemical effects in host extracts be demonstrated. PSII inactivation was correlated with increased levels of reactive oxygen species (ROS), degradation of pigments in peridinin-chl a protein, and chloroplast disruption, indicating that the FIS was under physiological collapse. The specific mechanism(s) causing Montipora-associated Symbiodinium to fail during the isolation methods used here remain unknown. However, these data indicate that care should be taken when using FIS to represent Symbiodinium in hospite, especially when comparing different species of corals.

Bentazone and bromoxynil induce H + and H 2O 2 accumulation, and inhibit photosynthetic O 2 evolution in Synechococcouselongatus PCC7942

Using a unicellular cyanobacterium, Synechococcous elongatus PCC7942, we have shown that cytosolic acidification, O 2 −; H 2O 2 production and photosystem II-inactivation are the causes of cell death from bentazone/bromoxynil incubations. Butyric acid evoked solely pH lowering response and yet inhibited PS II activity indicating that herbicide-caused acidification is sufficient to kill the cyanobacterial cells, but other factors like excess H 2O 2 production due to an imbalance in the peroxide sequestration machinery might be contributory. While the activities of superoxide dismutase and pyrogallol peroxidase increased consequent to herbicide incubations and displayed oligomeric states with mobility shift, catalase and glutathione peroxidase though present remained insensitive.

Photoprotection of reaction centres in photosynthetic organisms: mechanisms of thermal energy dissipation in desiccated thalli of the lichen Lobaria pulmonaria

*The photobionts of lichens have previously been shown to reversibly inactivate their photosystem II (PSII) upon desiccation, presumably as a photoprotective mechanism. The mechanism and the consequences of this process have been investigated in the green algal lichen Lobaria pulmonaria. *Lichen thalli were collected from a shaded and a sun-exposed site. The activation of PSII was followed by chlorophyll fluorescence measurements. *Inactivation of PSII, as indicated by the total loss of variable fluorescence, was accompanied by a strong decrease of basal fluorescence (F(0)). Sun-grown thalli, as well as thalli exposed to low irradiance during drying, showed a larger reduction of F(0) than shade-grown thalli or thalli desiccated in the dark. Desiccation increased phototolerance, which was positively correlated to enhanced quenching of F(0). Quenching of F(0) could be reversed by heating, and could be inhibited by glutaraldehyde but not by the uncoupler nigericin. *Activation of energy dissipation, apparent as F(0) quenching, is proposed to be based on an alteration in the conformation of a pigment protein complex. This permits thermal energy dissipation and gives considerable flexibility to photoprotection. Zeaxanthin formation apparently did not contribute to the enhancement of photoprotection by desiccation in the light. Light-induced absorbance changes indicated the involvement of chlorophyll and carotenoid cation radicals.

Acclimation of photosystem II to high temperature in a suspension culture of soybean (Glycine max) cells requires proteins that are associated with the thylakoid membrane

In a study of the responses of photosystem II (PSII) to high temperature in suspension-cultured cells of soybean (Glycine max L. Merr.), we found that high temperatures inactivated PSII via two distinct pathways. Inactivation of PSII by moderately high temperatures, such as 41 degrees C, was reversed upon transfer of cells to 25 degrees C. The recovery of PSII required light, but not the synthesis of proteins de novo. By contrast, temperatures higher than 45 degrees C inactivated PSII irreversibly. An increase in the growth temperature from 25 to 35 degrees C resulted in an upward shift of 3 degrees C in the profile of the heat-induced inactivation of PSII, which indicated that the thermal stability of PSII had been enhanced. This acclimative response was reflected by the properties of isolated thylakoid membranes: PSII in thylakoid membranes from cells that had been grown at 35 degrees C exhibited greater thermal stability than that from cells grown at 25 degrees C. Disruption of the vesicular structure of thylakoid membranes with 0.05% Triton X-100 decreased the thermal stability of PSII to a similar level in both types of thylakoid membrane. Proteins released by Triton X-100 from thylakoid membranes from cells grown at 35 degrees C were able to increase the thermal stability of Triton-treated thylakoid membranes. These observations suggest that proteins that are associated with thylakoid membranes might be involved in the enhancement of the thermal stability of PSII.