Photoinhibitory Treatment Research Articles

Photoinhibition and recovery of primary photosynthesis in Antarctic and subantarctic lichens. Analysis of interspecific differences

This study meticulously investigates the dynamics of photoinhibition and the mechanisms of primary photosynthetic activity recovery in lichens found in Antarctica and the sub-Antarctic regions. Advanced methodologies were utilised, such as Kautsky's kinetic analysis and the OJIP test. The study carefully details the response of various lichen species to intense light stress, outlining both immediate effects and subsequent recovery processes. Our findings reveal that these lichens employ a range of adaptive strategies, specific to each species, to mitigate the effects of photoinhibition, thereby emphasizing their remarkable resilience and ecological importance in harsh environments. Notably, the investigation reveals the sophisticated interplay between inherent photoprotective mechanisms and the ecological adaptations that enable these lichens to thrive under such harsh conditions. The study not only advances our knowledge of plant physiology under stress but also enriches our insights into the survival strategies of terrestrial organisms facing global environmental changes. Three types of photoinhibitory treatments differing in their duration and strength were applied to 7 lichen species from Antarctica and South America (Isla Navarino). The lichens responded with a decrease in photosynthetic processes in photosystem II (FV/FM and ΦPSII declined), although they showed almost complete recovery in the following 5 h. This was attributed to the activation of photoprotective mechanisms, non-photochemical quenching (NPQ) in particular, during photoinhibitory treatments. Chlorophyll fluorescence parameters derived from slow Kautsky kinetics were correlated with those derived from the OJIP curve. Our study presents data that supports the conclusion of significant photoresistance of the studied lichen species in the hydrated state to photoinhibition induced by high doses of photosynthetically active radiation (PAR).

Resistance of Primary Photosynthesis to Photoinhibition in Antarctic Lichen Xanthoria elegans: Photoprotective Mechanisms Activated during a Short Period of High Light Stress.

The Antarctic lichen, Xanthoria elegans, in its hydrated state has several physiological mechanisms to cope with high light effects on the photosynthetic processes of its photobionts. We aim to investigate the changes in primary photochemical processes of photosystem II in response to a short-term photoinhibitory treatment. Several chlorophyll a fluorescence techniques: (1) slow Kautsky kinetics supplemented with quenching mechanism analysis; (2) light response curves of photosynthetic electron transport (ETR); and (3) response curves of non-photochemical quenching (NPQ) were used in order to evaluate the phenomenon of photoinhibition of photosynthesis and its consequent recovery. Our findings suggest that X. elegans copes well with short-term high light (HL) stress due to effective photoprotective mechanisms that are activated during the photoinhibitory treatment. The investigations of quenching mechanisms revealed that photoinhibitory quenching (qIt) was a major non-photochemical quenching in HL-treated X. elegans; qIt relaxed rapidly and returned to pre-photoinhibition levels after a 120 min recovery. We conclude that the Antarctic lichen species X. elegans exhibits a high degree of photoinhibition resistance and effective non-photochemical quenching mechanisms. This photoprotective mechanism may help it survive even repeated periods of high light during the early austral summer season, when lichens are moist and physiologically active.

Effects of High Irradiance and Low Water Temperature on Photoinhibition and Repair of Photosystems in Marimo (Aegagropila linnaei) in Lake Akan, Japan.

The green alga Aegagropila linnaei often forms spherical aggregates called "marimo" in Lake Akan in Japan. In winter, marimo are exposed to low water temperatures at 1-4 °C but protected from strong sunlight by ice coverage, which may disappear due to global warming. In this study, photoinhibition in marimo was examined at 2 °C using chlorophyll fluorescence and 830 nm absorption. Filamentous cells of A. linnaei dissected from marimo were exposed to strong light at 2 °C. Photosystem II (PSII) was markedly photoinhibited, while photosystem I was unaffected. When the cells with PSII damaged by the 4 h treatment were subsequently illuminated with moderate repair light at 2 °C, the maximal efficiency of PSII was recovered to the level before photoinhibition. However, after the longer photoinhibitory treatments, PSII efficiency did not recover by the repair light. When the cells were exposed to simulated diurnal light for 12 h per day, which was more ecological, the cells died within a few days. Our results showed new findings of the PSII repair at 2 °C and serious damage at the cellular level from prolonged high-light treatments. Further, we provided a clue to what may happen to marimo in Lake Akan in the near future.

Plants acclimate to Photosystem I photoinhibition by readjusting the photosynthetic machinery.

Photosynthetic light reactions require strict regulation under dynamic environmental conditions. Still, depending on environmental constraints, photoinhibition of Photosystem (PSII) or PSI occurs frequently. Repair of photodamaged PSI, in sharp contrast to that of PSII, is extremely slow and leads to a functional imbalance between the photosystems. Slow PSI recovery prompted us to take advantage of the PSI‐specific photoinhibition treatment and investigate whether the imbalance between functional PSII and PSI leads to acclimation of photosynthesis to PSI‐limited conditions, either by short‐term or long‐term acclimation mechanisms as tested immediately after the photoinhibition treatment or after 24 h recovery in growth conditions, respectively. Short‐term acclimation mechanisms were induced directly upon inhibition, including thylakoid protein phosphorylation that redirects excitation energy to PSI as well as changes in the feedback regulation of photosynthesis, which relaxed photosynthetic control and excitation energy quenching. Longer‐term acclimation comprised reprogramming of the stromal redox system and an increase in ATP synthase and Cytochrome b6f abundance. Acclimation to PSI‐limited conditions restored the CO2 assimilation capacity of plants without major PSI repair. Response to PSI inhibition demonstrates that plants efficiently acclimate to changes occurring in the photosynthetic apparatus, which is likely a crucial component in plant acclimation to adverse environmental conditions.

Resistance of Antarctic moss Sanionia uncinata to photoinhibition: chlorophyll fluorescence analysis of samples from the western and eastern coasts of the Antarctic Peninsula.

Interspecific differences in sensitivity of the Antarctic moss Sanionia uncinata from King George Island (KGI) and James Ross Island (JRI) to photoinhibitory treatment were studied in laboratory conditions using chlorophyll fluorescence techniques. Slow (Kautsky) and fast (OJIP) kinetics were used for the measurements. Samples were exposed to a short-term (60min) photoinhibitory treatment (PIT, 2000μmol·m-2 ·s-1 PAR). The photoinhibitory treatment (PIT) led to photoinhibition which was indicated by the decrease in FV /FM and ΦPSII in KGI but not in JRI samples. However, this decrease was small and full recovery was reached 90min after PIT termination. Non-photochemical quenching (NPQ) was activated during the PIT, and rapidly relaxed during recovery. Early stages of photoinhibition showed a drop in FV /FM and ΦPSII to minimum values within the first 10s of the PIT, with their subsequent increase apparent within fast (0-5min PIT) and slow (5-50min PIT) phases of adjustment. The PIT caused a decrease in the performance index (Pi_Abs), photosynthetic electron transport per reaction centre (RC) (ET0 /RC). The PIT induced an increase in thermal dissipation per RC (DI0 /RC), effectivity of thermal dissipation (Phi_D0 ), absorption per RC (ABS/RC) and trapping rate per RC (TR0 /RC). In conclusion, PIT led to only slight photoinhibition followed by fast recovery in S. uncinata from KGI and JRI, since FV /FM and ΦPSII returned to pre-photoinhibitory conditions. Therefore, S. uncinata might be considered resistant to photoinhibition even in the wet state. The KGI samples showed higher resistance to photoinhibition than the JRI samples.

Glycinebetaine mitigated the photoinhibition of photosystem II at high temperature in transgenic tomato plants.

Photosystem II (PSII), especially the D1 protein, is highly sensitive to the detrimental impact of heat stress. Photoinhibition always occurs when the rate of photodamage exceeds the rate of D1 protein repair. Here, genetically engineered codA-tomato with the capability to accumulate glycinebetaine (GB) was established. After photoinhibition treatment at high temperature, the transgenic lines displayed more thermotolerance to heat-induced photoinhibition than the control line. GB maintained high expression of LeFtsHs and LeDegs and degraded the damaged D1 protein in time. Meanwhile, the increased transcription of synthesis-related genes accelerated the de novo synthesis of D1 protein. Low ROS accumulation reduced the inhibition of D1 protein translation in the transgenic plants, thereby reducing protein damage. The increased D1 protein content and decreased phosphorylated D1 protein (pD1) in the transgenic plants compared with control plants imply that GB may minimize photodamage and maximize D1 protein stability. As D1 protein exhibits a high turnover, PSII maybe repaired rapidly and efficiently in transgenic plants under photoinhibition treatment at high temperature, with the resultant mitigation of photoinhibition of PSII.

Low temperature and high light dependent dynamic photoprotective strategies in Arabidopsis thaliana.

Arabidopsis thaliana has been recognized as a chilling tolerant species based on analysis of resistance to low temperature stress, however, the mechanisms involved in this tolerance are not yet clarified. The low temperature-induced effects are exacerbated when plants are exposed to low temperatures in the presence of high light irradiance but the experimental data on the impact of light intensity during cold stress and its influence during recovery from stress are rather limited. The main objective of this study was to re-examine the photosynthetic responses of A. thaliana plants to short term (6 days) low temperature stress (12/10°C) under optimal (150 μmol m-2 s-1 ) and high light (500 μmol m-2 s-1 ) intensity and the subsequent recovery from the stress. Simultaneous measurements of the in vivo and in vitro functional performance of both photosystem II (PSII) and photosystem I (PSI), as well as, net photosynthesis, low temperature (77 K) chlorophyll fluorescence and immunoblot analysis of the relative abundance of PSII and PSI reaction center proteins were used to evaluate the role of light in the development of possible protective mechanisms during low temperature stress and the consequent recovery from exposure to low temperature and different light intensities. The results presented clearly suggest that Arabidopsis plants can employ a number of highly dynamic photoprotective strategies depending on the light intensity. These strategies include one based on LHCII quenching and two other quenching mechanisms localized within the PSII and PSI reaction centers, which are all expressed to different extent depending on the severity of the photoinhibitory treatments under low temperature stress conditions.

EGY3: homologue of S2P protease located in chloroplasts

The EGY3 protein is a homologue of site-2 proteases, which are intramembrane zinc metalloproteases. EGY3 itself lacks proteolytic activity due to the absence of a zinc-binding motif. Plentiful evidence indicates that such intramembrane 'pseudoproteases' play significant roles in many diverse processes occurring within the cell. However, the physiological functions of EGY3, as well as its subcellular localization, remain unknown. The subcellular localization of EGY3 protein was investigated using Arabidopsis thaliana protoplasts transformed with EGY3-GFP fusion protein, and immunoblot experiments using the total leaf protein extract, as well as highly purified chloroplasts and fractions of stroma, envelope and thylakoid membrane proteins. The physiological role of EGY3 was studied using two A.thaliana mutant lines devoid of EGY3 protein. Chlorophyll a fluorescence measurement was performed and the egy3 mutant sensitivity to photoinhibition was investigated. Additionally, the abundance of thylakoid membrane complexes was established using blue native gel electrophoresis. We present experimental evidence for thylakoid membrane localization of the EGY3 protein. We show that egy3 mutants display increased value of the non-photochemical quenching parameter and significantly slower recovery rate after photoinhibitory treatment. This was associated with a decrease in the level of proteases involved in photosystem II recovery, Deg1 and FtsH2/8.

PGR5 and NDH-1 systems do not function as protective electron acceptors but mitigate the consequences of PSI inhibition

Avoidance of photoinhibition at photosystem (PS)I is based on synchronized function of PSII, PSI, Cytochrome b6f and stromal electron acceptors. Here, we used a special light regime, PSI photoinhibition treatment (PIT), in order to specifically inhibit PSI by accumulating excess electrons at the photosystem (Tikkanen and Grebe, 2018). In the analysis, Arabidopsis thaliana WT was compared to the pgr5 and ndho mutants, deficient in one of the two main cyclic electron transfer pathways described to function as protective alternative electron acceptors of PSI. The aim was to investigate whether the PGR5 (pgr5) and the type I NADH dehydrogenase (NDH-1) (ndho) systems protect PSI from excess electron stress and whether they help plants to cope with the consequences of PSI photoinhibition. First, our data reveals that neither PGR5 nor NDH-1 system protects PSI from a sudden burst of electrons. This strongly suggests that these systems in Arabidopsis thaliana do not function as direct acceptors of electrons delivered from PSII to PSI – contrasting with the flavodiiron proteins that were found to make Physcomitrella patens PSI resistant to the PIT. Second, it is demonstrated that under light-limiting conditions, the electron transfer rate at PSII is linearly dependent on the amount of functional PSI in all genotypes, while under excess light, the PGR5-dependent control of electron flow at the Cytochrome b6f complex overrides the effect of PSI inhibition. Finally, the PIT is shown to increase the amount of PGR5 and NDH-1 as well as of PTOX, suggesting that they mitigate further damage to PSI after photoinhibition rather than protect against it.

Rate Constants of PSII Photoinhibition and its Repair, and PSII Fluorescence Parameters in Field Plants in Relation to their Growth Light Environments.

The extent of photoinhibition of PSII is determined by a balance between the rate of photodamage to PSII and that of repair of the damaged PSII. It has already been indicated that the rate constants of photodamage (kpi) and repair (krec) of the leaves differ depending on their growth light environment. However, there are no studies using plants in the field. We examined these rate constants and fluorescence parameters of several field-grown plants to determine inter-relationships between these values and the growth environment. The kpi values were strongly related to the excess energy, EY, of the puddle model and non-regulated energy dissipation, Y(NO), of the lake model, both multiplied by the photosynthetically active photon flux density (PPFD) level during the photoinhibitory treatment. In contrast, the krec values corrected against in situ air temperature were very strongly related to the daily PPFD level. The plants from the fields showed higher NPQ than the chamber-grown plants, probably because these field plants acclimated to stronger lightflecks than the averaged growth PPFD. Comparing chamber-grown plants and the field plants, we showed that kpi is determined by the incident light level and the photosynthetic capacities such as in situ rate of PSII electron transport and non-photochemical quenching (NPQ) [e.g. Y(NO)×PPFD] and that krec is mostly determined by the growth light and temperature levels.

Influence of in vitro growth conditions on the photosynthesis and survival of Castanea sativa plantlets during ex vitro transfer

Adequate in vitro micro-environments are crucial to induce life compatible leaf development. Key morphological and physiological traits are needed to allow ex vitro survival. We study, how in vitro light and ventilation affect physiological performance and survival of ex vitro Castanea sativa plantlets. In vitro treatments consisted of two irradiances of 50 and 150 µmol m−2 s−1 in ventilated vessels (VL50 and VL150, respectively), compared to traditional cultures at 50 µmol m−2 s−1 in non-ventilated vessels (NVL50). After the exposure to each condition a photoinhibitory treatment (PhT) was also applied to study whether the above in vitro conditions exerted photoprotection and facilitated the recovery of C. sativa during sudden ex vitro transfer. During rooting, a decrease in net photosynthesis (Psat), electron transport rate (ETRII) and maximal efficiency of PSII (F v /F m ) were observed. Transpiration rates (E) decreased, concomitantly with a rise in water use efficiency (WUE), mainly in microplants originating from ventilated treatments (V). Throughout this stage, the PhT was lethal for all in vitro treatments. During acclimation, the number and leaf size increased principally in plantlets originating from V treatments. These microplants were also able to recover their ETR and F v /F m . Initially, the PhT produced a drastic drop in Fv/Fm of plantlets in all treatments however they did show a tendency to recover. Transferring plantlets to the greenhouse produced a decrease in the Psat in all treatments; however, over time Psat increased reaching values of 3.2 and 5.3 μmol CO2 m−2 s−1 in microplants originating from VL50 and VL150, respectively. Transpiration rate were similar in all treatments and remained at levels of about 0.9 mmol H2O m−2 s−1; thus, WUE increased significantly, reaching values of almost 3.8 µmol CO2/mmol H2O in microplants originating from VL150. After the PhT, all of the plantlet’s recovery capacity increased concomitantly with their dynamic heat dissipation and their de-epoxidation capacity. Our results suggest that managing in vitro conditions can improve plantlets photosynthetic performance in early stages after ex vitro transfer, playing a key role in the ameliorating the transfer stress.

Photoinhibition of photosynthesis in Antarctic lichen Usnea antarctica. II. Analysis of non-photochemical quenching mechanisms activated by low to medium light doses

The paper focus sensitivity of an Antarctic lichen Usnea antarctica to photoinhibition studied under controlled laboratory conditions. Main emphasis was given to the analysis of quenching mechanisms, i.e. deexcitation pathways of absorbed light energy exploited in non-photochemical processes. Thalli of U. antarctica were collected at the James Ross Island, Antarctica (57°52´57´´ W, 63°48´02´´ S) and transferred in dry state to the Czech Republic. After rewetting in a laboratory, they were exposed to medium light intensities (300, 600 and 1000 mmol m-2 s-1 of photosynthetically active radiation) for 6 h. Before and during photoinhibitory treatments, chlorophyll fluorescence parameters, photoinhibitory (qI), state 1-2 transition (qT), and energy-dependent quenching (qE) in particular were measured to evaluate dose- and time-dependent changes in these parameters. The results showed that among the components forming non-photochemical quenching (qN), qI contributes to the largest extent to qN, while qE and qT contribute less. This finding differs from our earlier studies made in a short term-, and high light-treated U. antarctica that found qE together with qI is the most important part of non-photochemical quenching. Possible explanation is that photoinhibition in PS II in U. ant-arctica, when induced by low to medium light, activates qE to only limited extend and for a relatively short time (tens of minutes). With prolonged high light treatment lasting several hours, qE tends to be reduced to the values close to zero and qI then forms a major part of qN.

Xanthophyll Cycle Pool Size and Composition in Several Cosmarium Strains (Zygnematophyceae, Streptophyta) are Related to their Geographic Distribution Patterns

The photosynthetic behaviour and composition of photosynthetic pigments of four Cosmarium strains collected from different geographic areas were examined under moderate and photoinhibitory white light by means of PAM fluorometry and high-performance liquid chromatography. Generally, all of the Cosmarium strains displayed the photosynthetic performance and the composition of xanthophyll cycle pigments corresponding to that of high-light adapted plants and algae, when grown under the standard laboratory conditions. However, photoinhibitory treatments provoked several strain- and species-specific characteristics despite the long-term cultivation in laboratory conditions. The typical arctic taxon, C. crenatum var. boldtianum, displayed an incomplete violaxanthin cycle yielding an accumulation of antheraxanthin during high light stress, which is considered as an adaptation to occasional high irradiances in the polar zone due to the albedo. So far, the violaxanthin/antheraxanthin turnover was known only in some prasinophycean algae. Antheraxanthin actively participated in the heat dissipation from PSII centres in C. crenatum, as concluded from a significant positive correlation between non-photochemical quenching (NPQ) and the quantity of antheraxanthin. In contrast, all the other Cosmarium strains displayed a complete violaxanthin de-epoxidase action during the high light treatments, as judged from the relatively high production of zeaxanthin which participated in thermal dissipation of excess energy.

Inhibition of Calvin–Benson cycle suppresses the repair of photosystem II in Symbiodinium: implications for coral bleaching

Inhibition of Calvin–Benson cycle (CBC) activity by thermal stress has been hypothesized to cause photoinhibition of photosystem II (PSII) in zooxanthellae of reef-building corals and consequently lead to bleaching. This study tests whether the interruption of CBC by glycolaldehyde (GA) leads to photoinhibition and subsequent coral bleaching in Stylophora pistillata. When S. pistillata was incubated with GA, the O2 evolution rate declined in a dose-dependent manner and the extent of photoinhibition, reflected by a decreased maximum quantum yield of PSII (F v/F m), was enhanced. The effect of GA on photoinhibition was similar to that of chloramphenicol (CAP), an inhibitor of protein synthesis in chloroplasts. When S. pistillata was incubated in weak light following a high-light-induced photoinhibitory treatment, the recovery of PSII from photoinhibition was suppressed in a similar manner to both GA- and CAP-treated samples. After incubation in moderate light at 26°C, S. pistillata showed a bleaching response only in presence of GA. These results suggest that coral bleaching-like responses are caused by interruption of the CBC activity in S. pistillata and are associated with accelerated photoinhibition through suppression of the protein synthesis-dependent repair of PSII but not to an increase in photodamage to PSII.

Light energy management in micropropagated plants of Castanea sativa, effects of photoinhibition

The limited development of photoprotective mechanisms, specifically heat dissipation capacity, found in micropropagated plants may be the result of low xanthophyll cycle pigment content and reduced de-epoxidation capacity making them highly susceptible to photodamage. The effects of gradual or sudden increase of light on Castanea sativa in vitro cultured and during their ex vitro transference was evaluated. The results were compared with those determined in nursery-grown plants. In vitro plants responded poorly to gradual increase in irradiance, exhibiting a low electron transport rate (ETR) agreeing with low non-photochemical quenching (NPQ) and a limited de-epoxidation capacity, not synthesizing detectable amounts of zeaxanthin (Z). Regarding a sudden increase in light (photoinhibition treatment, PhT); post-PhT as in vitro as well nursery plants showed a significant decrease in their maximal efficiency of PSII (Fv/Fm), but in vitro the decrease was very drastic (around 0.2) different from that observed in nursery (around 0.69). In vitro, NPQ was mainly determined by the slow relaxing component, NPQs (80.8%), concomitant with a pronounced decrease of D1 protein post-PhT, and a lack of de-epoxidation capacity. During ex vitro transfer, PhT lead to death of some plants, specifically during root induction. The photoprotective mechanisms were activated over time in ex vitro conditions, indicating that micropropagated Castanea sativa display a potential for light acclimation, adjusting their photosynthetic apparatus to the ambient growth irradiance. Understanding the mechanisms that micropropagated plants deployed and how they face high light intensity events, will allow us to search for strategies to improve performance to possible light fluctuations that normally occur in ex vitro conditions during plant acclimation.

Cold-acclimation limits low temperature induced photoinhibition by promoting a higher photochemical quantum yield and a more effective PSII restoration in darkness in the Antarctic rather than the Andean ecotype of Colobanthus quitensis Kunt Bartl (Cariophyllaceae)

BackgroundEcotypes of Colobanthus quitensis Kunt Bartl (Cariophyllaceae) from Andes Mountains and Maritime Antarctic grow under contrasting photoinhibitory conditions, reaching differential cold tolerance upon cold acclimation. Photoinhibition depends on the extent of photodamage and recovery capability. We propose that cold acclimation increases resistance to low-temperature-induced photoinhibition, limiting photodamage and promoting recovery under cold. Therefore, the Antarctic ecotype (cold hardiest) should be less photoinhibited and have better recovery from low-temperature-induced photoinhibition than the Andean ecotype. Both ecotypes were exposed to cold induced photoinhibitory treatment (PhT). Photoinhibition and recovery of photosystem II (PSII) was followed by fluorescence, CO2 exchange, and immunoblotting analyses.ResultsThe same reduction (25%) in maximum PSII efficiency (Fv/Fm) was observed in both cold-acclimated (CA) and non-acclimated (NA) plants under PhT. A full recovery was observed in CA plants of both ecotypes under dark conditions, but CA Antarctic plants recover faster than the Andean ecotype.Under PhT, CA plants maintain their quantum yield of PSII, while NA plants reduced it strongly (50% and 73% for Andean and Antarctic plants respectively). Cold acclimation induced the maintenance of PsaA and Cyt b6/f and reduced a 41% the excitation pressure in Antarctic plants, exhibiting the lowest level under PhT. xCold acclimation decreased significantly NPQs in both ecotypes, and reduced chlorophylls and D1 degradation in Andean plants under PhT.NA and CA plants were able to fully restore their normal photosynthesis, while CA Antarctic plants reached 50% higher photosynthetic rates after recovery, which was associated to electron fluxes maintenance under photoinhibitory conditions.ConclusionsCold acclimation has a greater importance on the recovery process than on limiting photodamage. Cold acclimation determined the kinetic and extent of recovery process under darkness in both C. quitensis ecotypes. The greater recovery of PSII at low temperature in the Antarctic ecotype was related with its ability to maintain PsaA, Cyt b6/f and D1 protein after photoinhibitory conditions. This is probably due to either a higher stability of these polypeptides or to the maintenance of their turnover upon cold acclimation. In both cases, it is associated to the maintenance of electron drainage from the intersystem pool, which maintains QA more oxidized and may allow the synthesis of ATP and NADPH necessaries for the regeneration of ribulose 1,5-bisphosphate in the Calvin Cycle. This could be a key factor for C. quitensis success under the harsh conditions and the short growing period in the Maritime Antarctic.

Quenching partitioning through light-modulated chlorophyll fluorescence: A quantitative analysis to assess the fate of the absorbed light in the field

Plants use a small part of the total absorbed light energy for net carboxylation, while the remaining amount is dissipated via alternative pathways involving thermal processes, fluorescence and non-carboxylation photochemistry in order to limit the formation of reactive oxygen species (ROS) and other photooxidative risks. The commonly used analysis of the Photosystem II (PSII) fluorescence signals gives qualitative information about absorbed light energy management by plants, but it is difficult to appreciate the relative contribution of each pathway in energy partitioning. This study reports the application of quenching partitioning through a chlorophyll fluorescence approach performed on peach leaves subjected to three different light intensities for four durations of exposure in absence of recovery from photo-damage. This methodology was compared with the P 700 redox kinetic method for determining the functional PSII fraction in leaves. In the absence of recovery processes the active PSII concentration decayed with an increase in photon exposure (the product of irradiance and the time of exposure), following an exponential pattern according to the reciprocity law. The photoprotective thermal dissipation ( Φ NPQ ) was proportional to irradiance up to 30 min of photoinhibitory treatment. Afterwards Φ NPQ was limited by the increasing competition for the absorbed energy re-emitted by the inactive PSII ( Φ NF ). Φ NF increased with the photon exposure dissipating up to 70% of the total incoming energy. The energy funnelled to photochemistry ( Φ PSII ) decreased with increasing exposure time or intensity, becoming zero after 120 min of photoinhibitory treatment at the maximum irradiance (2100 μmol photon m −2 s −1). The relation between the fraction of energy dissipated by the inactive PSII (derived from the quenching partitioning) and the inactive PSII fraction (measured with the P 700 redox kinetic method) was linear. The quenching partitioning through light-modulated chlorophyll fluorescence is a useful tool to analyse plant energy management and gives also a reasonable estimation of the active PSII fraction. This methodology can easily be used in the field as measurements are rapid, non-destructive and detection devices are portable.

Evidence for light-independent and steeply decreasing PSII efficiency along twig depth in four tree species

Recent reports have indicated a considerably inactivated PSII in twig cortices, in spite of the low light transmittance of overlying periderms. Corresponding information for more deeply located and less illuminated tissues like xylem rays and pith are lacking. In this investigation we aimed to characterize the efficiency of PSII and its light sensitivity along twig depth, in conjunction with the prevailing light quantity and quality. To that aim, optical methods (spectral reflectance and transmittance, chlorophyll fluorescence imaging, low temperature fluorescence spectra) and photoinhibitory treatments were applied in cut twig sections of four tree species, while corresponding leaves served as controls. Compared to leaves, twig tissues displayed lower chlorophyll (Chl) levels and dark-adapted PSII efficiency, with strong decreasing gradients towards the twig center. The low PSII efficiencies in the inner stem were not an artifact due to an actinic effect of measuring beam or to an enhanced contribution of PSI fluorescence. In fact, the PSII/PSI ratios in cortices were higher and those in the xylem rays similar to that of leaves. Inner twig tissues were quite resistant to photoinhibitory treatments, tolerating irradiation levels several-fold higher than those encountered in their microenvironment. Moreover, the extent of high light tolerance was similar in naturally exposed and shaded twig sides. The results indicate an increasing, inherent and light-independent inactivation of PSII along twig depth. The findings are discussed on the basis of a recently proposed model for photosynthetic electron flow in twigs, taking into account the specific atmospheric and light microenvironment as well as the possible metabolic needs of such bulky organs.

Differential turnover of the photosystem II reaction centre D1 protein in mesophyll and bundle sheath chloroplasts of maize

Photoinhibition is caused by an imbalance between the rates of the damage and repair cycle of photosystem II D1 protein in thylakoid membranes. The PSII repair processes include (i) disassembly of damaged PSII–LHCII supercomplexes and PSII core dimers into monomers, (ii) migration of the PSII monomers to the stroma regions of thylakoid membranes, (iii) dephosphorylation of the CP43, D1 and D2 subunits, (iv) degradation of damaged D1 protein, and (v) co-translational insertion of the newly synthesized D1 polypeptide and reassembly of functional PSII complex. Here, we studied the D1 turnover cycle in maize mesophyll and bundle sheath chloroplasts using a protein synthesis inhibitor, lincomycin. In both types of maize chloroplasts, PSII was found as the PSII–LHCII supercomplex, dimer and monomer. The PSII core and the LHCII proteins were phosphorylated in both types of chloroplasts in a light-dependent manner. The rate constants for photoinhibition measured for lincomycin-treated leaves were comparable to those reported for C3 plants, suggesting that the kinetics of the PSII photodamage is similar in C3 and C4 species. During the photoinhibitory treatment the D1 protein was dephosphorylated in both types of chloroplasts but it was rapidly degraded only in the bundle sheath chloroplasts. In mesophyll chloroplasts, PSII monomers accumulated and little degradation of D1 protein was observed. We postulate that the low content of the Deg1 enzyme observed in mesophyll chloroplasts isolated from moderate light grown maize may retard the D1 repair processes in this type of plastids.

Different thermal sensitivity of the repair of photodamaged photosynthetic machinery in cultured Symbiodinium species

Coral bleaching caused by heat stress is accompanied by photoinhibition, which occurs under conditions where the rate of photodamage to photosystem II (PSII) exceeds the rate of its repair, in the symbiotic algae (Symbiodinium spp.) within corals. However, the mechanism of heat stress-induced photoinhibition in Symbiodinium still remains poorly understood. In the present work, we have investigated the effect of elevated temperature on the processes associated with the repair of photodamaged PSII in cultured Symbiodinium (OTcH-1 and CS-73). Severe photoinhibition was observed at temperature exceeding 32 degrees C in Symbiodinium CS-73 cells grown at 25-34 degrees C but not in cultures of the more thermally tolerant Symbiodinium OTcH-1. After photoinhibition treatment by strong light, photodamaged PSII was repaired close to initial levels under low light at 25 degrees C in both OTcH-1 and CS-73. However, the repair was strongly inhibited by increased temperature exceeding 31 degrees C in CS-73 but only weakly in OTcH-1. We found that inhibition of the repair process in CS-73 is attributed to impairment of both protein synthesis-dependent and -independent repair processes and is at least partially caused by suppression of the de novo synthesis of thylakoid membrane proteins and impairment of the generation of DeltapH across the thylakoid membrane, respectively. Our results suggest that acceleration of photoinhibition by moderate heat stress is attributed primarily to inhibition of the repair of photodamaged PSII and that the photoinhibition sensitivity of Symbiodinium to heat stress is determined by the thermal sensitivity of the PSII repair processes.