Photoinhibitory Light Research Articles

RESISTANCE TO PHYTOPHTHORA1 promotes cytochrome b559 formation during early photosystem II biogenesis in Arabidopsis.

As an essential intrinsic component of photosystem II (PSII) in all oxygenic photosynthetic organisms, heme-bridged heterodimer cytochrome b559 (Cyt b559) plays critical roles in the protection and assembly of PSII. However, the underlying mechanisms of Cyt b559 assembly are largely unclear. Here, we characterized the Arabidopsis (Arabidopsis thaliana) rph1 (resistance to Phytophthora1) mutant, which was previously shown to be susceptible to the oomycete pathogen Phytophthora brassicae. Loss of RPH1 leads to a drastic reduction in PSII accumulation, which can be primarily attributed to the defective formation of Cyt b559. Spectroscopic analyses showed that the heme level in PSII supercomplexes isolated from rph1 is significantly reduced, suggesting that RPH1 facilitates proper heme assembly in Cyt b559. Due to the loss of RPH1-mediated processes, a covalently bound PsbE-PsbF heterodimer is formed during the biogenesis of PSII. In addition, rph1 is highly photosensitive and accumulates elevated levels of reactive oxygen species under photoinhibitory-light conditions. RPH1 is a conserved intrinsic thylakoid protein present in green algae and terrestrial plants, but absent in Synechocystis, and it directly interacts with the subunits of Cyt b559. Thus, our data demonstrate that RPH1 represents a chloroplast acquisition specifically promoting the efficient assembly of Cyt b559, probably by mediating proper heme insertion into the apo-Cyt b559 during the initial phase of PSII biogenesis.

Psb28 protein is indispensable for stable accumulation of PSII core complexes in Arabidopsis.

Enhancing the efficiency of photosynthesis represents a promising strategy to improve crop yields, with keeping the steady state of PSII being key to determining the photosynthetic performance. However, the mechanisms whereby the stability of PSII is maintained in oxygenic organisms remain to be explored. Here, we report that the Psb28 protein functions in regulating the homeostasis of PSII under different light conditions in Arabidopsis thaliana. The psb28 mutant is much smaller than the wild-type plants under normal growth light, which is due to its significantly reduced PSII activity. Similar defects were seen under low light and became more pronounced under photoinhibitory light. Notably, the amounts of PSII core complexes and core subunits are specifically decreased in psb28, whereas the abundance of other representative components of photosynthetic complexes remains largely unaltered. Although the PSII activity of psb28 was severely reduced when subjected to high light, its recovery from photoinactivation was not affected. By contrast, the degradation of PSII core protein subunits is dramatically accelerated in the presence of lincomycin. These results indicate that psb28 is defective in the photoprotection of PSII, which is consistent with the observation that the overall NPQ is much lower in psb28 compared to the wild type. Moreover, the Psb28 protein is associated with PSII core complexes and interacts mainly with the CP47 subunit of PSII core. Taken together, these findings reveal an important role for Psb28 in the protection and stabilization of PSII core in response to changes in light environments.

Exposed anthocyanic leaves of Prunus cerasifera are special shade leaves with high resistance to blue light but low resistance to red light against photoinhibition of photosynthesis.

The photoprotective role of foliar anthocyanins has long been ambiguous: exacerbating, being indifferent to or ameliorating the photoinhibition of photosynthesis. The photoinhibitory light spectrum and failure to separate photo-resistance from repair, as well as the different methods used to quantify the photo-susceptibility of the photosystems, could lead to such a discrepancy. We selected two congeneric deciduous shrubs, Prunus cerasifera with anthocyanic leaves and Prunus triloba with green leaves, grown under identical growth conditions in an open field. The photo-susceptibilities of photosystem II (PSII) and photosystem I (PSI) to red light and blue light, in the presence of lincomycin (to block the repair), of exposed leaves were quantified by a non-intrusive P700+ signal from PSI. Leaf absorption, pigments, gas exchange and Chl a fluorescence were also measured. The content of anthocyanins in red leaves (P. cerasifera) was >13 times greater than that in green leaves (P. triloba). With no difference in maximum quantum efficiency of PSII photochemistry (Fv/Fm) and apparent CO2 quantum yield (AQY) in red light, anthocyanic leaves (P. cerasifera) showed some shade-acclimated suites, including lower Chl a/b ratio, lower photosynthesis rate, lower stomatal conductance and lower PSII/PSI ratio (on an arbitrary scale), compared with green leaves (P. triloba). In the absence of repair of PSII, anthocyanic leaves (P. cerasifera) showed a rate coefficient of PSII photoinactivation (ki) that was 1.8 times higher than that of green leaves (P. triloba) under red light, but significantly lower (-18 %) under blue light. PSI of both types of leaves was not photoinactivated under blue or red light. In the absence of repair, anthocyanic leaves exhibited an exacerbation of PSII photoinactivation under red light and a mitigation under blue light, which can partially reconcile the existing controversy in terms of the photoprotection by anthocyanins. Overall, the results demonstrate that appropriate methodology applied to test the photoprotection hypothesis of anthocyanins is critical.

The Passage of H2O2 from Chloroplasts to Their Associated Nucleus during Retrograde Signalling: Reflections on the Role of the Nuclear Envelope.

The response of chloroplasts to adverse environmental cues, principally increases in light intensity, stimulates chloroplast-to-nucleus retrograde signalling, which leads to the induction of immediate protective responses and longer-term acclimation. Hydrogen peroxide (H2O2), generated during photosynthesis, is proposed to both initiate and transduce a retrograde signal in response to photoinhibitory light intensities. Signalling specificity achieved by chloroplast-sourced H2O2 for signal transduction may be dependent upon the oft-observed close association of a proportion of these organelles with the nucleus. In this review, we consider more precisely the nature of the close association between a chloroplast appressed to the nucleus and the requirement for H2O2 to cross both the double membranes of the chloroplast and nuclear envelopes. Of particular relevance is that the endoplasmic reticulum (ER) has close physical contact with chloroplasts and is contiguous with the nuclear envelope. Therefore, the perinuclear space, which transducing H2O2 molecules would have to cross, may have an oxidising environment the same as the ER lumen. Based on studies in animal cells, the ER lumen may be a significant source of H2O2 in plant cells arising from the oxidative folding of proteins. If this is the case, then there is potential for the ER lumen/perinuclear space to be an important location to modify chloroplast-to-nucleus H2O2 signal transduction and thereby introduce modulation of it by additional different environmental cues. These would include for example, heat stress and pathogen infection, which induce the unfolded protein response characterised by an increased H2O2 level in the ER lumen.

Fast Diffusion of the Unassembled PetC1-GFP Protein in the Cyanobacterial Thylakoid Membrane.

Biological membranes were originally described as a fluid mosaic with uniform distribution of proteins and lipids. Later, heterogeneous membrane areas were found in many membrane systems including cyanobacterial thylakoids. In fact, cyanobacterial pigment–protein complexes (photosystems, phycobilisomes) form a heterogeneous mosaic of thylakoid membrane microdomains (MDs) restricting protein mobility. The trafficking of membrane proteins is one of the key factors for long-term survival under stress conditions, for instance during exposure to photoinhibitory light conditions. However, the mobility of unbound ‘free’ proteins in thylakoid membrane is poorly characterized. In this work, we assessed the maximal diffusional ability of a small, unbound thylakoid membrane protein by semi-single molecule FCS (fluorescence correlation spectroscopy) method in the cyanobacterium Synechocystis sp. PCC6803. We utilized a GFP-tagged variant of the cytochrome b6f subunit PetC1 (PetC1-GFP), which was not assembled in the b6f complex due to the presence of the tag. Subsequent FCS measurements have identified a very fast diffusion of the PetC1-GFP protein in the thylakoid membrane (D = 0.14 − 2.95 µm2s−1). This means that the mobility of PetC1-GFP was comparable with that of free lipids and was 50–500 times higher in comparison to the mobility of proteins (e.g., IsiA, LHCII—light-harvesting complexes of PSII) naturally associated with larger thylakoid membrane complexes like photosystems. Our results thus demonstrate the ability of free thylakoid-membrane proteins to move very fast, revealing the crucial role of protein–protein interactions in the mobility restrictions for large thylakoid protein complexes.

PHOTOPROTECTIIVE STRATEGIES OF UNICELULAR RED ALGA RHODELLA VIOLACEA DURING ACCLIMATION TO STRONG LIGHT

Red algae contain in their photosynthetic machinery water-soluble antenna complexes - phycobilisomes (PBSs) attached to thylakoid membranes to transfer excitation energy to photosystems. Strong light absorbed by the PBSs triggers a fast formation of transthylakoid ΔpH that follows the non-photochemical quenching of chlorophyll (Chl) fluorescence. The ΔpH build-up seems to be essential for photoprotecting the photosynthetic apparatus in the absence of xanthophyll cycle common to higher plants. However, the photoprotective mechanisms of red algae are not studied in details yet. 
 We present here our research of the Chl fluorescence quenching in unicellular red algae Rhodella violacea and its correlation with the ΔpH gradient being formed. The relation of this phenomenon to photoprotection of photosystem 2 (PS 2) in the normal and high light-acclimated Rhodella cells is also examined. 
 Under the photoinhibitory conditions (white light of 2000-3000 μE/m2s), the ΔpH-dependent Chl fluorescence quenching was found to delay the kinetics of PS 2 photoinhibition. The uncouplers like nigericin and NH4Cl are known to break down ΔpH gradient, lead to the dissipation of Chl fluorescence quenching followed by enhancing the PS 2 photoinhibition rate. The same effect showed far-red (FR) light consuming transthylakoid ΔpH. ATPase inhibitor, DCCD, having no impact on ΔpH didn’t influence PS 2 photoinhibition as well this implies the photoprotection to be fulfilled by the proton gradient rather than by ATP synthesis. 
 Long-term acclimation of Rhodella cells to higher irradiances (500-1000 μE/m2s) results in a partial loss of the periphery phycoerythrin-containing subunits by PBSs. The light-acclimated cultures display a higher resistance to the photoinhibitory light than the non-acclimated ones. This could be explained by diminishing the energy transfer from the reduced PBSs to PS 2 as well as light screening by the secondary carotenoids synthesized during light exposure. 
 Data on low-temperature (77K) fluorescence allow to evaluate the molecular mechanisms of light-induced Chl fluorescence suppression in Rhodella cells and its recovery in darkness.

Effects of thinning and reshaping on canopy micro-domain environment and leaf physiological characteristics in dense Fuji apple orchard on Loess Plateau of eastern Gansu, China

We examined the effects of thinning and reshaping on the canopy micro-domain environment, leaves micro-structure, physiological characteristics and photosynthetic capacity of 16 year-old Fuji apple dense orchards by measuring canopy relative light intensity, temperature, relative humidity, leaf chlorophyll content, micro-structure, and photosynthetic fluorescence. After thinning and reshaping, relative light intensity and temperature of tree canopy were significantly improved, and the distribution was balanced. Effective light intensity (>30%) was 57% higher than that of control (thinning and reshaping, CK), and temperature increased by 1.1 ℃. Due to the improvement of relative light intensity and temperature in the canopy, leaf chlorophyll, leaf thickness, and palisade tissue thickness of thinning trees were increased by 8.7%, 5.4%, and 9.2%, respectively. Net photosynthetic rate, transpiration rate and stomatal conductance of the leaves were also significantly increased, being 12.6%, 17.1% and 7.3% higher than CK, respectively. Leaf photosynthesis capacity of both treatments was limited by non-stomatal factors. After thinning and reshaping, Fm (maximum fluorescence) and qN(non-photochemical quenching coefficient) of PSⅡ in leaves were increased by 1.5% and 2.1%. Leaves did not suffer strong light photoinhibition, with ABS/RC (unit reaction center absorbs light energy), ETo/RC (energy used for electron transfer captured by unit reaction center) and TRo/RC (energy captured by the unit reaction center to reduce QA) of leaves being significantly improved. The physiological characteristics of leaves were closely related to light and temperature environment. After thinning, light and temperature of the orchard canopy were improved, which promoted leaves growth and development, improved foliar micro-structure and photosynthetic efficiency. Thinning and reshaping were suitable strategies for the adjustment and optimization of Fuji orchard density in Loess Plateau of eastern Gansu.

Response of the thylakoid proteome of Synechocystis sp. PCC 6803 to photohinibitory intensities of orange-red light

Photoautotrophic growth of Synechocystis sp. PCC 6803 in a flat-panel photobioreactor, run in turbidostat mode under increasing intensities of orange-red light (636 nm), showed a maximal growth rate (0.12 h−1) at 300 μmolphotons m−2 s−1, whereas first signs of photoinhibition were detected above 800 μmolphotons m−2 s−1. To investigate the dynamic modulation of the thylakoid proteome in response to photoinhibitory light intensities, quantitative proteomics analyses by SWATH mass spectrometry were performed by comparing thylakoid membranes extracted from Synechocystis grown under low-intensity illumination (i.e. 50 μmolphotons m−2 s−1) with samples isolated from cells subjected to photoinhibitory light regimes (800, 950 and 1460 μmolphotons m−2 s−1). We identified and quantified 126 proteins with altered abundance in all three photoinhibitory illumination regimes.These data reveal the strategies by which Synechocystis responds to photoinibitory growth irradiances of orange-red light. The accumulation of core proteins of Photosystem II and reduction of oxygen-evolving-complex subunits in photoinhibited cells revealed a different turnover and repair rates of the integral and extrinsic Photosystem II subunits with variation of light intensity. Furthermore, Synechocystis displayed a differentiated response to photoinhibitory regimes also regarding Photosystem I: the amount of PsaD, PsaE, PsaJ and PsaM subunits decreased, while there was an increased abundance of the PsaA, PsaB, Psak2 and PsaL proteins. Photoinhibition with 636 nm light also elicited an increased capacity for cyclic electron transport, a lowering of the amount of phycobilisomes and an increase of the orange carotenoid protein content, all presumably as a photoprotective mechanism against the generation of reactive oxygen species.

Different responses of Echinochloa crus-galli and Echinochloa colona to Bisbyric-sodium (Nomineetm)

Echinochloa crus-galli and Echinochloa colona mostly grown as weeds in different economic crops and cause losses in their yield. Flooding and Crop rotation ca not suppress these weeds. Herbicides are widely applied to control barnyard grasses. Bisbyric-sodium was applied at FRD (Field Recommended Dose) on both species, the results of preliminary experiment indicated that E. crus-galli was susceptible to bispyribac-sodium but E. colona was resistance. The Confirmation experiment for E. colona resistance indicated that herbicide effect on the biomass of plants which caused reduction in fresh and dry weight. The photosynthetic pigments were affected by herbicide, which decreased by increasing the herbicide concentration. While anthocyanins content were increased to protect the leaves from the stress of photoinhibitory light fluxes by absorbing the excess photons. The soluble and insoluble sugar content was increased by increasing the concentration of herbicide. The transcript level of ALS in E. colona was modified by different concentrations of herbicide. The present study indicated that ALS expression level was responsive to herbicide treatment. This suggests that ALS is a target site of bispyric-sodium. But 50% FRD can be considered as sub-lethal dose which upregulated the expression of ALS by 4 folds.

Chlorophyll fluorescence and excitation energy dissipation of pot-grown Phyllostachys rivalis leaves after long-term flooding.

To determine the physiological and biochemical responses and adaptive mechanisms of Phyllostachys rivalis to long-term soil flooding, two-year-old potted seedlings of Ph. rivalis were subjected to different flooding depths. The flooding treatment was set with water levels of 5 cm (TreatmentⅠ) and 10 cm (TreatmentⅡ) higher than the soil surface and normal water supply (ck). Then the chlorophyll fluorescence parameters in leaves were determined after continuous flooding for 30, 90, 180, 270, and 360 d. Responses to the continuous flooding stress for light energy absorption and transformation, energy transfer and distribution, reaction center activity, and excitation energy dissipation in leaves of three leaves per seedlings and three seedlings were measured and analyzed. A one-way analysis of variance with duncan's test was conducted at a significant level of 0.05. Results showed that the chlorophyll fluorescence parameters varied with different flooding levels. When flooding for 30 d and 90 d, minimal fluorescence (Fo), maximum fluorescence (Fm), photochemical maximum efficiency of PS Ⅱ (Fv/Fm), photochemical quenching coefficient (qP), and electron transport rate(RET) of Treatment Ⅰ were not significantly different (P > 0.05) compared with ck; however, Treatment Ⅱ when flooding for 90 d compared with ck was significantly increased for Fo while significantly decreased for Fm, Fv/Fm and RET (P 0.05). The amount of absorbed light in photochemistry (P) and excess energy (E) increased first and then decreased (P < 0.05), but the energy of dissipation of the antenna heat dissipation (D) observed were opposite. Taken together, Ph. rivalis could maintain a relatively high RET, qP, and P in the early stages of flooding, and enhance qN to regulate their energy metabolism, dissipate excess light energy via heat dissipation, thereby alleviating the light photoinhibition and photooxidation; however, continuous long-term soil flooding could damage photosystem Ⅱ (PS Ⅱ) even though there was minimal adverse effect on the functional center of PS Ⅱ (P < 0.05). It can be inferred that Ph. rivalis can tolerate a short term flooding which facilitate its possible application in plantation restoration of riparion zone.

A chloroplast thylakoid lumen protein is required for proper photosynthetic acclimation of plants under fluctuating light environments

Despite our increasingly sophisticated understanding of mechanisms ensuring efficient photosynthesis under laboratory-controlled light conditions, less is known about the regulation of photosynthesis under fluctuating light. This is important because-in nature-photosynthetic organisms experience rapid and extreme changes in sunlight, potentially causing deleterious effects on photosynthetic efficiency and productivity. Here we report that the chloroplast thylakoid lumenal protein MAINTENANCE OF PHOTOSYSTEM II UNDER HIGH LIGHT 2 (MPH2; encoded by At4g02530) is required for growth acclimation of Arabidopsis thaliana plants under controlled photoinhibitory light and fluctuating light environments. Evidence is presented that mph2 mutant light stress susceptibility results from a defect in photosystem II (PSII) repair, and our results are consistent with the hypothesis that MPH2 is involved in disassembling monomeric complexes during regeneration of dimeric functional PSII supercomplexes. Moreover, mph2-and previously characterized PSII repair-defective mutants-exhibited reduced growth under fluctuating light conditions, while PSII photoprotection-impaired mutants did not. These findings suggest that repair is not only required for PSII maintenance under static high-irradiance light conditions but is also a regulatory mechanism facilitating photosynthetic adaptation under fluctuating light environments. This work has implications for improvement of agricultural plant productivity through engineering PSII repair.

Effects of periodic photoinhibitory light exposure on physiology and productivity of Arabidopsis plants grown under low light

This work examined the long-term effects of periodic high light stress on photosynthesis, morphology, and productivity of low-light-acclimated Arabidopsis plants. Significant photoinhibition of Arabidopsis seedlings grown under low light (100 μmol photons m-2 s-1) was observed at the beginning of the high light treatment (three times a day for 30 min at 1800 μmol photons m-2 s-1). However, after 2 weeks of treatment, similar photosynthesis yields (Fv/Fm) to those of control plants were attained. The daily levels of photochemical quenching measured in the dark (qPd) indicated that the plants recovered from photoinhibition within several hours once transferred back to low light conditions, with complete recovery being achieved overnight. Acclimation to high light stress resulted in the modification of the number, structure, and position of chloroplasts, and an increase in the average chlorophyll a/b ratio. During ontogenesis, high-light-exposed plants had lower total leaf areas but higher above-ground biomass. This was attributed to the consumption of starch for stem and seed production. Moreover, periodic high light exposure brought forward the reproductive phase and resulted in higher seed yields compared with control plants grown under low light. The responses to periodic high light exposure of mature Arabidopsis plants were similar to those of seedlings but had higher light tolerance.

Regulation of Intertidal Microphytobenthos Photosynthesis Over a Diel Emersion Period Is Strongly Affected by Diatom Migration Patterns.

Changes in biomass and photosynthesis of a diatom-dominated microphytobenthos (MPB) intertidal community were studied over a diel emersion period using a combination of O2 and scalar irradiance microprofiling, variable chlorophyll (Chl) fluorescence, and pigment analysis. The MPB biomass in the photic zone (0–0.5 mm) of the sediment exposed to low irradiance (150 μmol photons m-2 s-1) showed a >2-fold increase during the first hours of the emersion period, reaching >0.2 mg Chl a cm-3. Concentrations of Chl a started to decrease half-way through the emersion period, almost 2 h before tidal inundation. Similarly, O2 concentrations and volumetric gross photosynthesis in the photic zone increased during the first half of the emersion period and then decreased toward the timing of incoming tide/darkness. The results suggest that intertidal MPB community-level photosynthesis is mainly controlled by changes in the productive biomass of the photic zone determined by cell migration. A diel pattern in the photosynthesis vs. irradiance parameters α (photosynthetic efficiency at limiting irradiance) and ETRmax (photosynthetic capacity at saturating irradiance) was also observed, suggesting photoacclimation of MPB. Under high light exposure (2000 μmol photons m-2 s-1), lower α, ETRmax and sediment O2 concentrations were observed when cell migration was inhibited with the diatom motility inhibitor latrunculin A (Lat A), showing that migration is also used by MPB to maximize photosynthesis by reducing exposure to potentially photoinhibitory light levels. A higher de-epoxidation state in sediment treated with Lat A indicates that the involvement of the xanthophyll cycle in physiological photoprotection is more relevant in MPB when cells are inhibited from migrating. In the studied diatom-dominated MPB intertidal community, cell migration seems to be the key factor regulating photosynthesis over a diel emersion period and upon changes in light exposure.

Elucidation of Photoprotective Mechanisms of PSI Against Fluctuating Light photoinhibition.

It has been claimed that the cyclic electron flow around PSI (CEF-PSI) plays an important role in protection of PSI against fluctuating light photoinhibition. However, the photoprotective mechanism of PSI is not fully elucidated. Here, we examined the mechanism, using two CEF-PSI mutants of Arabidopsis thaliana, and antimycin A, an inhibitor of the PGR5 (proton gradient regulation 5)-mediated CEF-PSI. Dark-adapted leaves in these plants were illuminated in fluctuating light alternating between high light at 1,200 µmol m-2 s-1 and low light at 30 µmol m-2 s-1 every 2 min, and PSI and PSII parameters were simultaneously measured for 160 min with 830 nm absorption and Chl fluorescence, respectively. When CEF-PSI, especially PGR5-mediated CEF-PSI, did not operate, the acceptor-side limitation of PSI, Y(NA), increased stepwise, leading to marked PSI photoinhibition. The deficiency of CFE-PSI decreased not only the electron transport rate through PSI but also the donor-side limitation of PSI, Y(ND), in high light phases. These results showed that the large Y(ND), observed only when CEF-PSI operated, contributed to suppression of PSI photoinhibition. Taken together with our previous report that high Y(NA) was alleviated by the enhancement of CEF-PSI, a model for the protective mechanisms of PSI is proposed. In this model, both alleviation of Y(NA) and acceleration of Y(ND) are indispensable, and for realization of such a situation, regulation of the electron flows, especially the PGR5-mediated CEF-PSI, plays a key role. It is important for effective protection to regulate the balance of Y(ND) and Y(NA) through CEF-PSI.

MPH1 is a thylakoid membrane protein involved in protecting photosystem II from photodamage in land plants

Photosystem II (PSII) is highly susceptible to photoinhibition caused by environmental stimuli such as high light; therefore plants have evolved multifaceted mechanisms to efficiently protect PSII from photodamage. We previously published data suggesting that Maintenance of PSII under High light 1 (MPH1, encoded by AT5G07020), a PSII-associated proline-rich protein found in land plants, participates in the maintenance of normal PSII activity under photoinhibitory stress. Here we provide additional evidence for the role of MPH1 in protecting PSII against photooxidative damage. Two Arabidopsis thaliana mutants lacking a functional MPH1 gene suffer from severe photoinhibition relative to the wild-type plants under high irradiance light. The mph1 mutants exhibit significantly decreased PSII quantum yield and electron transport rate after exposure to photoinhibitory light. The mutants also display drastically elevated photodamage to PSII reaction center proteins after high-light treatment. These data add further evidence that MPH1 is involved in PSII photoprotection in Arabidopsis. MPH1 homologs are found across phylogenetically diverse land plants but are not detected in algae or prokaryotes. Taken together, these results suggest that MPH1 protein began to play a role in protecting PSII against excess light following the transition from aquatic to terrestrial conditions.

A land plant-specific thylakoid membrane protein contributes to photosystem II maintenance in Arabidopsis thaliana.

The structure and function of photosystem II (PSII) are highly susceptible to photo-oxidative damage induced by high-fluence or fluctuating light. However, many of the mechanistic details of how PSII homeostasis is maintained under photoinhibitory light remain to be determined. We describe an analysis of the Arabidopsis thaliana gene At5g07020, which encodes an unannotated integral thylakoid membrane protein. Loss of the protein causes altered PSII function under high-irradiance light, and hence it is named 'Maintenance of PSII under High light 1' (MPH1). The MPH1 protein co-purifies with PSII core complexes and co-immunoprecipitates core proteins. Consistent with a role in PSII structure, PSII complexes (supercomplexes, dimers and monomers) of the mph1 mutant are less stable in plants subjected to photoinhibitory light. Accumulation of PSII core proteins is compromised under these conditions in the presence of translational inhibitors. This is consistent with the hypothesis that the mutant has enhanced PSII protein damage rather than defective repair. These data are consistent with the distribution of the MPH1 protein in grana and stroma thylakoids, and its interaction with PSII core complexes. Taken together, these results strongly suggest a role for MPH1 in the protection and/or stabilization of PSII under high-light stress in land plants.

Antioxidant System Against Active Oxygen Species in Cyanobacterium Aphanothece stagnina: Response to Excess Light Under Cadmium Stress

The aim of the present study was to investigate the effect of photoinhibitory light induced changes under cadmium stress in electron transport activity, active oxygen species (AOS) generation, oxidative damage, photorespiration, enzymatic and non-enzymatic (low molecular weight) antioxidants in cyanobacterium, Aphanothece stagnina. The photosynthetic pigment contents enhanced progressively with increasing light intensity (20–120 μmol photons m−2 s−1); however, different light intensities together with cadmium (4 μM) decreased the pigment contents. Accumulation of Cd in cyanobacterial cells increased appreciably under photoinhibitory light (1,000 μmol photons m−2 s−1) treatment. Photoinhibitory light inhibited photosynthetic electron transport activity. In the presence of elevated level (50–100 μM) of Cd the inhibition became stronger showing enhanced effect on whole chain activity and photosystem II (PS II) with the least effect on photosystem I. Under photoinhibitory light treatment, the level of oxidants like hydrogen peroxide and superoxide radical increased substantially with simultaneous increase in lipid peroxidation and decrease in membrane intactness. The presence of Cd further enhanced such effects. Under this condition, considerable increase in photorespiration (loss of 14CO2) and content of non-enzymatic antioxidant like ascorbate, and appreciable suppression in the activity of enzymatic antioxidants like peroxidase, superoxide dismutase and catalase were noticed. These results indicate that photoinhibitory light alone and together with Cd strongly arrested electron flow through PS II and significantly inhibited the activity of enzymatic antioxidants. As consequence of this substantial rise in the level of AOS induced oxidative damage in A. stagnina.

Photoprotection by foliar anthocyanins mitigates effects of boron toxicity in sweet basil (Ocimum basilicum).

Boron (B) toxicity is an important agricultural problem in arid environments. Excess edaphic B compromises photosynthetic efficiency, limits growth and reduces crop yield. However, some purple-leafed cultivars of sweet basil (Ocimum basilicum) exhibit greater tolerance to high B concentrations than do green-leafed cultivars. We hypothesised that foliar anthocyanins protect basil leaf mesophyll from photo-oxidative stress when chloroplast function is compromised by B toxicity. Purple-leafed 'Red Rubin' and green-leafed 'Tigullio' cultivars, grown with high or negligible edaphic B, were given a photoinhibitory light treatment. Possible effects of photoabatement by anthocyanins were simulated by superimposing a purple polycarbonate filter on the green leaves. An ameliorative effect of light filtering on photosynthetic quantum yield and on photo-oxidative load was observed in B-stressed plants. In addition, when green protoplasts from both cultivars were treated with B and illuminated through a screen of anthocyanic protoplasts or a polycarbonate film which approximated cyanidin-3-O-glucoside optical properties, the degree of photoinhibition, hydrogen peroxide production, and malondialdehyde content were reduced. The data provide evidence that anthocyanins exert a photoprotective role in purple-leafed basil mesophyll cells, thereby contributing to improved tolerance to high B concentrations.

Simplifying data acquisition in plant canopies‐ Measurements of leaf angles with a cell phone

Summary Canopies are complex multilayered structures comprising individual plant crowns exposing a multifaceted surface area to sunlight. Foliage arrangement and properties are the main mediators of canopy functions. The leaves act as light traps whose exposure to sunlight varies with time of the day, date and latitude in a trade‐off between photosynthetic light harvesting and excessive or photoinhibitory light avoidance. To date, ecological research based upon leaf sampling has been limited by the available technology, with which data acquisition becomes labour intensive and time‐consuming, given the overwhelming number of leaves involved. In the present study, our goal involved developing a tool capable of measuring a sufficient number of leaves to enable analysis of leaf populations, tree crowns and canopies. We specifically tested whether a cell phone working as a 3D pointer could yield reliable, repeatable and valid leaf angle measurements with a simple gesture. We evaluated the accuracy of this method under controlled conditions, using a 3D digitizer, and we compared performance in the field with the methods commonly used. We presented an equation to estimate the potential proportion of the leaf exposed to direct sunlight (SAL) at any given time and compared the results with those obtained by means of a graphical method. We found a strong and highly significant correlation between the graphical methods and the equation presented. The calibration process showed a strong correlation between the results derived from the two methods with a mean relative difference below 10%. The mean relative difference in calculation of instantaneous exposure was below 5%. Our device performed equally well in diverse locations, in which we characterized over 700 leaves in a single day. The new method, involving the use of a cell phone, is much more effective than the traditional methods or digitizers when the goal is to scale up from leaf position to performance of leaf populations, tree crowns or canopies. Our methodology constitutes an affordable and valuable tool within which to frame a wide range of ecological hypotheses and to support canopy modelling approaches.

Selected coccal green algae are not affected by the humic substance Huminfeed® in term of growth or photosynthetic performance

Humic substances (HSs) have been shown to influence growth, photophysiology, and redox homeostasis in phototrophs. However, many ecological studies deliver controversial results indicating inhibitory or stimulating effects depending on the kind of phototrophic organism or type of HSs applied. Here we analyzed the effect of Huminfeed® (HF), a preparation from leonardite associated with lignite deposits, on growth and photosynthetic performance of three different coccal green algae. Concentrations of HF in the range of 2–20 mg l−1 dissolved organic carbon (DOC) did neither affect the growth rate nor the light-adapted photosynthetic electron transport. Even photoinhibitory light intensities of 1,600 μmol photons m−2 s−1, representing the tenfold of growth light intensity, did not result in a decline of the maximal photosynthetic rate in HF-treated algae. In HF-grown algae, a very subtle decrease by about 10% could be observed in thermoluminescent light emission, a sensitive method to detect changes in photosystem II (PSII) chemistry. However, thermoluminescence (TL) represents only 3% of the light-induced charge separated states in PSII. Hence, rather small changes will not have significant effects on overall photosynthetic performance and growth. Therefore, the physiological effect of HSs on freshwater phototrophs has to be revisited.