Photoprotective Mechanisms Research Articles

Current Knowledge on Mechanisms Preventing Photosynthesis Redox Imbalance in Plants.

Photosynthesis includes a set of redox reactions that are the source of reducing power and energy for the assimilation of inorganic carbon, nitrogen and sulphur, thus generating organic compounds, and oxygen, which supports life on Earth. As sessile organisms, plants have to face continuous changes in environmental conditions and need to adjust the photosynthetic electron transport to prevent the accumulation of damaging oxygen by-products. The balance between photosynthetic cyclic and linear electron flows allows for the maintenance of a proper NADPH/ATP ratio that is adapted to the plant’s needs. In addition, different mechanisms to dissipate excess energy operate in plants to protect and optimise photosynthesis under adverse conditions. Recent reports show an important role of redox-based dithiol–disulphide interchanges, mediated both by classical and atypical chloroplast thioredoxins (TRXs), in the control of these photoprotective mechanisms. Moreover, membrane-anchored TRX-like proteins, such as HCF164, which transfer electrons from stromal TRXs to the thylakoid lumen, play a key role in the regulation of lumenal targets depending on the stromal redox poise. Interestingly, not all photoprotective players were reported to be under the control of TRXs. In this review, we discuss recent findings regarding the mechanisms that allow an appropriate electron flux to avoid the detrimental consequences of photosynthesis redox imbalances.

Differential response of the photosynthetic machinery to dehydration in older and younger resurrection plants.

A group of vascular plants called homoiochlorophyllous resurrection plants evolved unique capabilities to protect their photosynthetic machinery against desiccation-induced damage. This study examined whether the ontogenetic status of the resurrection plant Craterostigma pumilum has an impact on how the plant responds to dehydration at the thylakoid membrane level to prepare cells for the desiccated state. Thus, younger plants (<4 months) were compared with their older (>6 months) counterparts. Ultrastructural analysis provided evidence that younger plants suppressed senescence-like programs that are realized in older plants. During dehydration, older plants degrade specific subunits of the photosynthetic apparatus such as the D1 subunit of PSII and subunits of the cytochrome b6f complex. The latter leads to a controlled down-regulation of linear electron transport. In contrast, younger plants increased photoprotective high-energy quenching mechanisms and maintained a high capability to replace damaged D1 subunits. It follows that depending on the ontogenetic state, either more degradation-based or more photoprotective mechanisms are employed during dehydration of Craterostigma pumilum.

Identification of sequence motifs in Lhcx proteins that confer qE-based photoprotection in the diatom Phaeodactylum tricornutum.

Photosynthetic organisms in nature often experience light fluctuations. While low light conditions limit the energy uptake by algae, light absorption exceeding the maximal rate of photosynthesis may go along with enhanced formation of potentially toxic reactive oxygen species. To preempt high light-induced photodamage, photosynthetic organisms evolved numerous photoprotective mechanisms. Among these, energy-dependent fluorescence quenching (qE) provides a rapid mechanism to dissipate thermally the excessively absorbed energy. Diatoms thrive in all aquatic environments and thus belong to the most important primary producers on earth. qE in diatoms is provided by a concerted action of Lhcx proteins and the xanthophyll cycle pigment diatoxanthin. While the exact Lhcx activation mechanism of diatom qE is unknown, two lumen-exposed acidic amino acids within Lhcx proteins were proposed to function as regulatory switches upon light-induced lumenal acidification. By introducing a modified Lhcx1 lacking these amino acids into a Phaeodactylum tricornutum Lhcx1-null qE knockout line, we demonstrate that qE is unaffected by these two amino acids. Based on sequence comparisons with Lhcx4, being incapable of providing qE, we perform domain swap experiments of Lhcx4 with Lhcx1 and identify two peptide motifs involved in conferring qE. Within one of these motifs, we identify a tryptophan residue with a major influence on qE establishment. This tryptophan residue is located in close proximity to the diadinoxanthin/diatoxanthin-binding site based on the recently revealed diatom Lhc crystal structure. Our findings provide a structural explanation for the intimate link of Lhcx and diatoxanthin in providing qE in diatoms.

Phenoplate: An innovative method for assessing interacting effects of temperature and light on non-photochemical quenching in microalgae under chemical stress

Rapid light curves are one of the most widely used methods for assessing the physiological state of photosynthetic organisms. While the method has been applied in a range of physiological studies over the last 20 years, little progress has been made in adapting it for the new age of multi-parametric phenotyping. In order to advance research that is aimed at evaluating the physiological impact of multiple factors, the Phenoplate was developed: a simultaneous assessment of temperature and light gradients. It was used to measure rapid light curves of three marine microalgae across a temperature gradient and altered phosphate availability. The results revealed that activation of photoprotective mechanisms occurred with high efficiency at lower temperatures, and relaxation of photoprotection was negatively impacted above a certain temperature threshold in Tetraselmis sp. It was observed that Thalassiosira pseudonana and Nannochloropsis oceanica exhibited two unique delayed non-photochemical quenching signatures: in combinations of low light with low temperature, and darkness with high temperature, respectively. These findings demonstrate that the Phenoplate approach can be used as a rapid and simple tool to gain insight into the photobiology of microalgae.

Durum wheat ideotypes in Mediterranean environments differing in water and temperature conditions

Ideotypic characteristics of durum wheat associated with higher yield under different water and temperature regimes were studied under Mediterranean conditions. Six semi-dwarf cultivars with contrasting agronomic performance were grown during two consecutive years under winter-planted rainfed and winter-planted support-irrigation conditions and a late-planting trial under support irrigation, at the INIA station of Colmenar de Oreja (Madrid). Different traits were assessed to inform on: water status, root performance, phenology, photosynthetic capacity, crop growth, grain yield and agronomic yield components. Under support irrigation and normal planting, genotypes with higher grain yield exhibited better water status (lower δ13C and canopy temperature), assimilation of more superficial water (higher δ18O), earlier heading and greater plant height and ear density. Under water-limited conditions (rainfed), the best genotypes also exhibited better water status (lower δ13C) and earlier heading, but higher specific root length with extraction of water from deeper soil layers (lower δ18O), more efficient N metabolism (higher δ15N and NBI) and consequently stronger growth (plant height and NDVI), and greater ear density and thousand grain weight. Under warmer conditions (late planting), the best genotypes also exhibited better water status (lower δ13C) and greater plant height and photoprotective mechanisms (higher flavonoid content and lower chlorophyll content). However, the strong differences in drought between consecutive years determined other specific ideotypic traits within each of the three growing conditions and the particular year. Our study suggests specific ideotypes when breeding durum wheat under different agronomic scenarios, but also stresses that interannual variation in water conditions, typical of Mediterranean conditions, should be taken into account.

Loss of carbonic anhydrase in the thylakoid lumen causes unusual moderate-light-induced rearrangement of the chloroplast in Chlamydomonas reinhardtii as a way of photosystem II photoprotection

Chlamydomonas reinhardtii cells have a single large cup-shaped chloroplast that can lose lobes under high light to prevent photodamage of the photosynthetic apparatus, including photosystem II (PSII). Here, under moderate light treatment, the development of the unusual morphology of the chloroplast is shown for mutant cia3, which is deficient in carbonic anhydrase (EC 4.2.1.1) CAH3 in the thylakoid lumen, while such light intensity is harmless for wild type (WT) cells for hours. Cia3 cells had more activated PSII photoprotective mechanisms and showed a tendency to shift in the balance of the PSII damage-repair cycle, whereas PSII retained the same photosynthetic efficiency as in the WT. These findings allow speculation about the unique PSII photoprotection strategy by rearranging the chloroplast in the absence of CAH3. CAH3, in turn, is suggested to be an important participant of the C. reinhardtii photosynthetic apparatus operation, functioning in close connection with PSII.

The impact of magnesium deficiency on photosynthesis and photoprotection in Spinacia oleracea

Limited magnesium (Mg) supply adversely affects photosynthesis. This is particularly related to the high demand for Mg of key enzymes in the chloroplast, such as the photosystems, the ATP synthase and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). The accepted critical Mg concentrations for yield and dry matter (DM) are 1.5–3.5 mg Mg g−1 DM. Earlier studies on Mg deficiency indicated that carbon fixation by Rubisco is severely affected in various plant species, whereas the impact of Mg scarcity on light reactions and photoprotective mechanisms is quite variable. The latter could be related to species-specific differences in the general high light-sensitivity of photosynthetic light reactions. To test this hypothesis, we studied the impact of Mg deficiency in spinach (Spinacia oleracea) plants, which are known to be rather high light resistant. S. oleracea seeds were grown hydroponically under four Mg treatments (1 (control), 0.05, 0.025 and 0.015 mM) and the impact of Mg deficiency on CO2 assimilation, photosynthetic light reactions and photoprotection was determined. Our results show that the photosynthetic efficiency and the overall light stress response were not altered under Mg deficiency, whereas the CO2 assimilation as well as leaf and root Mg concentrations were significantly reduced.

Photosynthetic physiological response of water-saving and drought-resistant rice to severe drought under wetting-drying alternation irrigation.

Water-saving and drought-resistant rice (WDR) is widely grown in central China in recent years. However, studies have not explored the interaction effect of WDR and irrigation regimes on drought-resistance capacities under severe drought at sensitive growth periods. A pot experiment was conducted using a WDR cultivar Hanyou73 (HY73) and traditional high-yielding and drought-sensitive cultivar Huiliangyou 898 (HLY898). Three irrigation regimes, including flooding irrigation (W1), mild wetting-drying alternation irrigation (W2), and severe wetting-drying alternation irrigation (W3), were applied before heading. At heading, severe drought with -50 KPa soil water potential was established for all treatments and cultivars. The findings showed that cultivar HY73 under W2 treatment had the highest yield, 1000-grain yield, filled grain, relative water content, and photosynthesis potential compared with the other combinations. The higher net photosynthetic rate (Pn ) was attributed to larger mesophyll conductance (gm ) in drought for cultivar HY73 under W2 treatment compared with that for cultivar HLY898 and the other water treatments. Enhanced photo-respiration rate may be an important photoprotection mechanism for achieving high Pn for cultivar HY73 coupled with W2 treatment than for other combinations in drought. The relative expression level of OsPIP1;1 gene was significantly down-regulated during drought in all cultivars and water regimes. But OsPIP1;2, OsPIP2;3, OsTIP2;2, and OsTIP3;1 genes were upregulated to alleviate the significant decrease in gs and gm under drought. These results suggest that WDR and mild wetting-drying alternation irrigation (W2) have significant interaction effects in improving photosynthetic production potential by maintaining higher gm under severe drought.

Epiphytic effects of Licmophora paradoxa on pigments of Pyropia yezoensis

Epiphytic diatoms usually cause adverse effects on photosynthesis of the host plants due to shading light or by interfering with the biochemical pathways. The present study investigated the epiphytic effects of the diatom Licmophora paradoxa on the pigments of red alga Pyropia yezoensis, such as the chlorophyll-a (Chl-a), Phycoerythrin (PE), Phycocyanin (PC), Allophycocyanin (APC) and carotenoid contents. The results showed that Chl-a was significantly decreased while other pigments such as PE, PC, APC and carotenoid contents were significantly increased in P. yezoensis due to the attachment of epiphytic diatom L. paradoxa. The present results indicated that epiphytic diatoms produced negative effects on the host PSII reaction center by reducing its main pigment Chl-a. Whereas, antenna pigments of Phycobiliprotein, of P. yezoensis such as PE, PC and APC were increased to capture more light energy supplying photosynthesis. The increase in carotenoid content under this epiphytic situation implied an enhancement of its assisted function in light-harvesting, photoprotection and stress-tolerance mechanism. The present findings contribute to well understanding the response mechanism of host macroalgae to epiphytic microalgae.

Uniform Water Potential Induced by Salt, Alkali, and Drought Stresses Has Different Impacts on the Seedling of Hordeum jubatum: From Growth, Photosynthesis, and Chlorophyll Fluorescence.

Hordeum jubatum is a halophyte ornamental plant wildly distributed in the Northeast of China, where the low water potential induced by various abiotic stresses is a major factor limiting plant growth and development. However, little is known about the comparative effects of salt, alkali, and drought stresses at uniform water potential on the plants. In the present study, the growth, gas exchange parameters, photosynthetic pigments, and chlorophyll fluorescence in the seedlings of H. jubatum under three low water potentials were measured. The results showed that the growth and photosynthetic parameters under these stresses were all decreased except for carotenoid (Car) with the increasing of stress concentration, and alkali stress caused the most damaging effects on the seedlings. The decreased net photosynthetic rate (Pn), stomatal conductance (Gs), and intercellular CO2 concentrations (Ci) values under salt stress were mainly attributed to stomatal factors, while non-stomatal factors were dominate under drought and alkali stresses. The reduced chlorophyll and slightly increased Car contents occurred under these stresses, and most significant changed under alkali stress. In addition, the maximum photochemical efficiency (Fv/Fm), actual photochemical efficiency (ΦPSII), and photochemical quenching coefficient (qP) under the stresses were all decreased, indicating that salt, alkali, and drought stresses all increased susceptibility of PSII to photoinhibition, reduced the photosynthetic activity by the declined absorption of light for photochemistry, and increased PSII active reaction centers. Moreover, the non-photochemical quenching coefficient (NPQ) of alkali stress was different from salt and drought stresses, showing that the high pH of alkali stress caused more damaging effects on the photoprotection mechanism depending on the xanthophyll cycle. The above results suggest that the H. jubatum has stronger tolerance of salt than drought and alkali stresses, and the negative effects of alkali stress on the growth and photosynthetic performance of this species was most serious.

PsbS-зависимые изменения в содержании липидов и фотосинтетических пигментов в растениях риса

Plants acclimate to fluctuations in light conditions by adjusting their photosynthetic apparatus. When the light intensity exceeds, an unbalanced excitation of the two photosystems occurs. It results in reduced photosynthetic efficiency. Photosystem II (PSII) is the most susceptible and dynamically regulated part of the light reactions in the thylakoid membrane. Non-photochemical quenching of chlorophyll fluorescence (NPQ) is one of the short-term photoprotective mechanisms, which consist of the number of components. The strongest NPQ component — qE is localized in the PSII antenna and induced in plants by lumen acidification, the activation of the pH sensor PsbS, and the conversion of the violaxanthin to zeaxanthin within the xanthophyll cycle. Here, I present data that characterizes the role of the PsbS protein in organization of PSII structural components in isolated PSII-enriched membranes. The preparations were isolated from wild-type (WT) and PsbS-less (PsbS-KO) mutant rice plant. Based on the obtained results, the PSII-enriched membranes from WT and PsbS-KO differ as in the level of lipids, also in carotenoids. I conclude that the PsbS-dependent changes in membrane fluidity in PsbS-KO mutant plants compensated with increased lipid level in mutant plants.

Thioredoxin-like protein CDSP32 alleviates Cd-induced photosynthetic inhibition in tobacco leaves by regulating cyclic electron flow and excess energy dissipation

Thioredoxin-like protein CDSP32 (Trx CDSP32), a thioredoxin-like (Trx-like) protein located in the chloroplast, can regulate photosynthesis and the redox state of plants under stress. In order to examine the role of Trx CDSP32 in the photosynthetic apparatus of plants exposed to cadmium (Cd), the effects of Trx CDSP32 on photosynthetic function and photoprotection in tobacco leaves under Cd exposure were studied using a proteomics approach with wild-type (WT) and Trx CDSP32 overexpression (OE) tobacco plants. Cd exposure reduced stomatal conductance, blocked PSII photosynthetic electron transport, and inhibited carbon assimilation. Increased water use efficiency (WUE), cyclic electron flow (CEF) of the proton gradient regulation 5 pathway (PGR5-CEF), and regulated energy dissipation [Y(NPQ)] are important mechanisms of Cd adaptation. However, CEF of the NAD(P)H dehydrogenase pathway (NDH-CEF) was inhibited by Cd exposure. Relative to control conditions, the expression levels of violaxanthin de-epoxidase (VDE) and photosystem II 22 kDa protein (PsbS) in OE leaves were significantly increased under Cd exposure, but those in WT leaves did not change significantly. Moreover, the expression of zeaxanthin epoxidase (ZE) under Cd exposure was significantly higher than that in WT leaves. Thus, Trx CDSP32 increased Y(NPQ) and alleviated PSII photoinhibition under Cd exposure. Trx CDSP32 not only increased PGR5-like protein 1A and 1B expression, but also alleviated the down-regulation of NAD(P)H-quinone oxidoreductase subunits induced by Cd exposure. Thus, Trx CDSP32 promotes CEF in Cd-exposed tobacco leaves. Thus, Trx CDSP32 alleviates the Cd-induced photoinhibition in tobacco leaves by regulating two photoprotective mechanisms: CEF and xanthophyll cycle-dependent energy dissipation.

Modelling Photoionisations in Tautomeric DNA Nucleobase Derivatives 7H-Adenine and 7H-Guanine: Ultrafast Decay and Photostability

The study of radiation effects in DNA is a multidisciplinary endeavour, connecting the physical, chemical and biological sciences. Despite being mostly filtered by the ozone layer, sunlight radiation is still expected to (photo)ionise DNA in sizeable yields, triggering an electron removal process and the formation of potentially reactive cationic species. In this manuscript, photoionisation decay channels of important DNA tautomeric derivatives, 7H-adenine and 7H-guanine, are characterised with accurate CASSCF/XMS-CASPT2 theoretical methods. These simulation techniques place the onset of ionisation for 7H-adenine and 7H-guanine on average at 8.98 and 8.43 eV, in line with recorded experimental evidence when available. Cationic excited state decays are analysed next, uncovering effective barrierless deactivation routes for both species that are expected to decay to their (cationic) ground state on ultrafast timescales. Conical intersection topographies reveal that these photoionisation processes are facilitated by sloped single-path crossings, known to foster photostability, and which are predicted to enable the (VUV) photo-protection mechanisms present in these DNA tautomeric species.

Light Induced Changes in Pigment and Lipid Profiles of Bryopsidales Algae

Bryopsidales (Chlorophyta) are cultured and consumed in several regions of the planet and are known for their high nutritional value and bioprospection potential, due to a high content of relevant polar lipids and polysaccharides. Among other characteristic features, these marine algae are known for possessing unique photosynthetic pigment-protein complexes and for the absence (in nearly all species investigated) of a functional xanthophyll cycle, a ubiquitous photoprotection mechanism present in most algae and plants. With the aim of characterizing the photophysiology of this atypical group of algae, we investigated the changes in pigment content and polar lipidome of two Bryopsidales species (Codium tomentosumandBryopsis plumosa) exposed for 7 days to low or high irradiance (20 vs. 1,000 μmol photons m–2s–1). Our results show that high light has a strong effect on the pigment composition, triggering the time-dependent accumulation of all-trans-neoxanthin (t-Neo) and violaxanthin (Viola). High light-acclimated macroalgae also displayed a shift in the characteristic polar lipidome, including a trend of accumulation of lyso-glycolipids, and highly unsaturated phospholipids and betaine lipids. We hypothesize that the observed shifts on the lipid composition could promote the interaction betweent-Neo and Viola with the siphonaxanthin–chlorophyll–protein complexes (SCP) of photosystem II (PSII) within the thylakoid membranes of the chloroplasts. Light induced changes in pigment and lipid composition could contribute to the fitness of Bryopsidales algae by reducing damages to the photosynthetic apparatus under increased irradiance.

Phlorotannin and Pigment Content of Native Canopy-Forming Sargassaceae Species Living in Intertidal Rockpools in Brittany (France): Any Relationship with Their Vertical Distribution and Phenology?

Five native Sargassaceae species from Brittany (France) living in rockpools were surveyed over time to investigate photoprotective strategies according to their tidal position. We gave evidences for the existence of a species distribution between pools along the shore, with the most dense and smallest individuals in the highest pools. Pigment contents were higher in lower pools, suggesting a photo-adaptive process by which the decreasing light irradiance toward the low shore was compensated by a high production of pigments to ensure efficient photosynthesis. Conversely, no xanthophyll cycle-related photoprotective mechanism was highlighted because high levels of zeaxanthin rarely occurred in the upper shore. Phlorotannins were not involved in photoprotection either; only some lower-shore species exhibited a seasonal trend in phlorotannin levels. The structural complexity of phlorotannins appears more to be a taxonomic than an ecological feature: Ericaria produced simple phloroglucinol while Cystoseira and Gongolaria species exhibited polymers. Consequently, tide pools could be considered as light-protected areas on the intertidal zone, in comparison with the exposed emerged substrata where photoprotective mechanisms are essential.

Light-harvesting complex stress-related proteins play crucial roles in the acclimation of Physcomitrella patens under fluctuating light conditions.

Photosynthetic organisms have evolved photoprotective mechanisms to acclimate to light intensity fluctuations in their natural growth environments. Photosystem (PS) II subunit S (PsbS) and light-harvesting complex (LHC) stress-related proteins (LhcSR) are essential for triggering photoprotection in vascular plants and green algae, respectively. The activity of both proteins is strongly enhanced in the moss Physcomitrella patens under high-light conditions. However, their role in regulating photosynthesis acclimation in P. patens under fluctuating light (FL) conditions is still unknown. Here, we compare the responses of wild-type (WT) P. patens and mutants lacking PsbS (psbs KO) or LhcSR1 and 2 (lhcsr KO) to FL conditions in which the low-light phases were periodically interrupted with high-light pulses. lhcsr KO mutant showed a strong reduction in growth with respect to WT and psbs KO under FL conditions. The lack of LhcSR not only decreased the level of non-photochemical quenching, resulting in an over-reduced plastoquinone pool, but also significantly increased the PSI acceptor limitation values with respect to WT and psbs KO under FL conditions. Moreover, in lhcsr KO mutant, the abundance of PSI core and PSI-LHCI complex decreased greatly under FL conditions compared with the WT and psbs KO. We proposed that LhcSR in P. patens play a crucial role in moss acclimation to dynamic light changes.

Molecular basis and regulatory mechanisms underlying fungal insecticides' resistance to solar ultraviolet irradiation.

Resistance to solar ultraviolet (UV) irradiation is crucial for field-persistent control efficacies of fungal formulations against arthropod pests, because their active ingredients are formulated conidia very sensitive to solar UV wavelengths. This review seeks to summarize advances in studies aiming to quantify, understand and improve conidial UV resistance. One focus of studies has been on the many sets of genes that have been revealed in the postgenomic era to contribute to or mediate UV resistance in the insect pathogens serving as main sources of fungal insecticides. Such genetic studies have unveiled the broad basis of UV-resistant molecules including cytosolic solutes, cell wall components, various antioxidant enzymes, and numerous effectors and signaling proteins, that function in developmental, biosynthetic and stress-responsive pathways. Another focus has been on the molecular basis and regulatory mechanisms underlying photorepair of UV-induced DNA lesions and photoreactivation of UV-impaired conidia. Studies have shed light upon a photoprotective mechanism depending on not only one or two photorepair-required photolyases, but also two white collar proteins and other partners that play similar or more important roles in photorepair via interactions with photolyases. Research hotspots are suggested to explore a regulatory network of fungal photoprotection and to improve the development and application strategies of UV-resistant fungal insecticides. © 2021 Society of Chemical Industry.

Diethyl sinapate-grafted cellulose nanocrystals as nature-inspired UV filters in cosmetic formulations.

Inspired by nature’s photoprotection mechanisms, we report an effective UV-blocking nanomaterial based on diethyl sinapate-grafted cellulose nanocrystals (CNC-DES). The colloidal stability and UV-blocking performance of CNC-DES in aqueous glycerol (a common humectant in petroleum-free cosmetic formulations) and in a commercially available moisturizing cream were studied. Grafting the water-insoluble DES onto CNCs renders it dispersible in these water-based formulations, thanks to the excellent water-dispersibility of CNC nanoparticles. Glycerol dispersions containing 0.1 to 1.5 wt% CNC-DES display very high UV-blocking activity owing to the anti-UV DES moieties anchored onto CNCs. A facial cream blended with 1.5 wt% CNC-DES exhibits an SPF of 5.03, which is higher than a commercially available sunscreen with the same active ingredient concentration (SPF = 3.84). DPPH radical scavenging assay also showed the antioxidant potential of CNC-DES, albeit coinciding with a significant reduction in antioxidant activity after grafting DES onto CNCs. Cytotoxicity measurements revealed the CNC-DES not to cause significant cytotoxicity to murine fibroblast cells after 24 h of exposure. Overall, CNC-DES exhibits strong anti-UV and antioxidant properties and is water-dispersible, biocompatible, non-greasy, and lightweight. This study demonstrates the exceptional potential of DES-grafted CNCs as nature-inspired UV filters in the next generation of cosmetic formulations, including those for sensitive skins.

Altering the exciton landscape by removal of specific chlorophylls in monomeric LHCII provides information on the sites of triplet formation and quenching by means of ODMR and EPR spectroscopies

The triplet states populated under illumination in the monomeric light-harvesting complex II (LHCII) were analyzed by EPR and Optically Detected Magnetic Resonance (ODMR) in order to fully characterize the perturbations introduced by site-directed mutations leading to the removal of key chlorophylls. We considered the A2 and A5 mutants, lacking Chls a612(a611) and Chl a603 respectively, since these Chls have been proposed as the sites of formation of triplet states which are subsequently quenched by the luteins. Chls a612 and Chl a603 belong to the two clusters determining the low energy exciton states in the complex. Their removal is expected to significantly alter the excitation energy transfer pathways. On the basis of the TR- and pulse EPR triplet spectra, the two symmetrically related pairs constituted by Chl a612/Lut620 and Chl a603/Lut621 were both possible candidate for triplet-triplet energy transfer (TTET). However, the ODMR results clearly show that only Lut620 is involved in triplet quenching. In the A5 mutant, the Chl a612/Lut620 pair retains this pivotal photoprotective role, while the A2 mutant was found to activate an alternative pathway involving the Chl a603/Lut621pair. These results shows that LHCII is characterized by a robust photoprotective mechanism, able to adapt to the removal of individual chromophores while maintaining a remarkable degree of Chl triplet quenching. Small amounts of unquenched Chl triplet states were also detected. The analysis of the results allowed us to assign the sites of “unquenched” chlorophyll triplets to Chl a610 and Chl a602.

Physiological and comparative transcriptome analysis of leaf response and physiological adaption to saline alkali stress across pH values in alfalfa (Medicago sativa)

Soil salinization is a critical factor limiting growth and causing physiological dysfunction in plants. The damage from alkaline salt in most plants is significantly greater than that from neutral salt. However, there is still a lack of research on the action mechanism by which saline alkali stress on plants under the same salt concentration across different pH values. The present study examined the effects of different pH values (7.0, 8.0, 9.0, and 10.0) under the same salt concentration (200 mmolL−1) on photosynthetic function, photoprotective mechanism, nitrogen metabolism, and osmotic regulation in alfalfa (Medicago sativa) leaves, including a transcriptomic analysis of changes in gene expression related to the above metabolic processes. The results showed that low pH saline alkali stress (pH 7.0 and 8.0) promoted chlorophyll synthesis in alfalfa leaves, and non-photochemical quenching (NPQ) and cyclic electron transfer (CEF) were promoted. There was no significant effect on plant growth or photochemical activity. The soluble sugar, proline, and soluble protein contents did not change significantly, and there was no obvious oxidative damage in alfalfa leaves. However, when pH increased to 9.0 and 10.0, KEGG enrichment analysis showed that photosynthesis (map00195) and nitrogen metabolism (map00910) were significantly enriched (P < 0.05), and PSII antenna protein coding genes were down-regulated under pH 9.0 and 10.0 treatments. The activities of PSII and PSI were decreased under high pH saline alkali stress, and the expression levels of the photosynthetic electron transporter-related genes PetA, PetB, petE, and petF were also significantly down-regulated. PSII was more sensitive to high pH saline alkali stress than PSI, and the PSII receptor side was more sensitive to high pH saline alkali stress than the PSII donor side. The activities of the oxygen-evolving complex (OEC) and PSI were significantly damaged only at pH 10.0. The activities of nitrate reductase (NR) and nitrite reductase (NiR), the expression levels of their genes, and the content of soluble protein were also decreased under pH 9.0 and 10.0 treatments. The inhibition of plant growth and oxidative damage to alfalfa leaves caused by high pH saline alkali stress were mainly related to the inhibition of photosynthesis (light energy absorption, electron transfer) and nitrogen metabolism (NO3− reduction). Under high pH saline alkali stress (pH 10.0), the photoprotection mechanisms such as CEF and NPQ were inhibited, which was also one of the important reasons for photoinhibition in alfalfa leaves. The accumulation of osmotic adjustment substances, such as soluble sugar and proline, was an important mechanism by which alfalfa physiologically adapted to high pH alkaline salt stress.