Xanthophyll Cycle Pigments Research Articles

Intervening dark periods negatively affect the photosynthetic performance of Chlamydomonas reinhardtii during growth under fluctuating high light.

The acclimation of the green algae Chlamydomoas reinhardtii to high light (HL) has been studied predominantly under continuous illumination of the cells. Here, we investigated the impact of fluctuating HL in alternation with either low light (LL) or darkness on photosynthetic performance and on photoprotective responses. Compared to intervening LL phases, dark phases led to (1) more pronounced reduction of the photosystem II quantum efficiency, (2) reduced degradation of the PsbS protein, (3) lower energy dissipation capacity and (4) an increased pool size of the xanthophyll cycle pigments. These characteristics indicate increased photo-oxidative stress when HL periods are interrupted by dark phases instead of LL phases. This overall trend was similar when comparing long (8 h) and short (30 min) HL phases being interrupted by long (16 h) and short (60 min) phases of dark or low light, respectively. Only the degradation of PsbS was clearly more efficient during long (16 h) LL phases when compared to short (60 min) LL phases.

Do red and yellow autumn leaves make use of different photoprotective strategies during autumn senescence?

Our goal was to determine whether anthocyanin-producing species (red) use different photoprotective strategies to cope with excess light during fall senescence compared with non-anthocyanin-producing species (yellow). In a previous study, we found that a yellow species retained the photoprotective PsbS protein in late autumn, while a red species did not. Specifically, we tested the hypothesis that red species make less use of zeaxanthin and PsbS-mediated thermal dissipation, as they rely on anthocyanins for photoprotection. We monitored four red (Acer ginnala, Rhus typhnia, Parenthocissus quinquefolia, Viburnum dentatum) and four yellow species (Acer negundo, Ostrya virginiana, Vitis riparia, Zanthoxylum americanum) throughout autumn senescence and analyzed pigments, protein content, and chlorophyll fluorescence. We found yellow species retained the PsbS protein at higher levels, and had higher dark retention of zeaxanthin in late autumn relative to red species. All species retained lutein and the pool of xanthophyll cycle pigments in higher amounts than other carotenoids in late autumn. Our data support the hypothesis that red species use anthocyanins as a photoprotective strategy during autumn senescence, and therefore make less use of PsbS and zeaxanthin-mediated thermal dissipation. We also found species-specific variation in the particular combination of photoprotective strategies used.

The Amount ofZeaxanthin Epoxidase But Not the Amount ofViolaxanthin De-Epoxidase Is a Critical Determinant ofZeaxanthin Accumulation inArabidopsis thaliana andNicotiana tabacum.

The generation of violaxanthin (Vx) de-epoxidase (VDE), photosystem II subunit S (PsbS) and zeaxanthin (Zx) epoxidase (ZEP) (VPZ) lines, which simultaneously overexpress VDE, PsbS and ZEP, has been successfully used to accelerate the kinetics of the induction and relaxation of non-photochemical quenching (NPQ). Here, we studied the impact of the overexpression of VDE and ZEP on the conversion of the xanthophyll cycle pigments in VPZ lines of Arabidopsis thaliana and Nicotiana tabacum. The protein amount of both VDE and ZEP was determined to be increased to about 3- to 5-fold levels of wild-type (WT) plants for both species. Compared to WT plants, the conversion of Vx to Zx, and hence VDE activity, was only marginally accelerated in VPZ lines, whereas the conversion of Zx to Vx, and thus ZEP activity, was strongly increased in VPZ lines. This indicates that the amount of ZEP but not the amount of VDE is a critical determinant of the equilibrium of the de-epoxidation state of xanthophyll cycle pigments under saturating light conditions. Comparing the two steps of epoxidation, particularly the second step (antheraxanthin to Vx) was found to be accelerated in VPZ lines, implying that the intermediate Ax is released into the membrane during epoxidation by ZEP.

Basking in the sun: how mosses photosynthesise and survive in Antarctica.

The Antarctic environment is extremely cold, windy and dry. Ozone depletion has resulted in increasing ultraviolet-B radiation, and increasing greenhouse gases and decreasing stratospheric ozone have altered Antarctica's climate. How do mosses thrive photosynthetically in this harsh environment? Antarctic mosses take advantage of microclimates where the combination of protection from wind, sufficient melt water, nutrients from seabirds and optimal sunlight provides both photosynthetic energy and sufficient warmth for efficient metabolism. The amount of sunlight presents a challenge: more light creates warmer canopies which are optimal for photosynthetic enzymes but can contain excess light energy that could damage the photochemical apparatus. Antarctic mosses thus exhibit strong photoprotective potential in the form of xanthophyll cycle pigments. Conversion to zeaxanthin is high when conditions are most extreme, especially when water content is low. Antarctic mosses also produce UV screening compounds which are maintained in cell walls in some species and appear to protect from DNA damage under elevated UV-B radiation. These plants thus survive in one of the harshest places on Earth by taking advantage of the best real estate to optimise their metabolism. But survival is precarious and it remains to be seen if these strategies will still work as the Antarctic climate changes.

Photoinhibition and Photoprotective Responses of a Brown Marine Macroalga Acclimated to Different Light and Nutrient Regimes.

Plants and brown algae avoid photoinhibition (decline in photosystem II efficiency, Fv/Fm) caused by excess light energy and oxidative stress through several photoprotective mechanisms, such as antioxidant xanthophyll production and heat dissipation. The heat dissipation can be measured as non-photochemical quenching (NPQ) and is strongly driven by de-epoxidation of xanthophyll cycle pigments (XCP). Although NPQ is known to increase under high light acclimation and nutrient-deficient conditions, a few studies have investigated the combined effects of the conditions on both NPQ and associated xanthophyll-to-chlorophyll (Chl) a ratio. The present study investigated the photosynthetic parameters of the brown alga Sargassum fusiforme acclimated to three irradiance levels combined with three nutrient levels. Elevated irradiance decreased Fv/Fm but increased NPQ, XCP/Chl a ratio, and fucoxanthin/Chl a ratio, suggesting the photoprotective role of antioxidant fucoxanthin in brown algae. Reduced nutrient availability increased NPQ but had no effect on the other variables, including XCP/Chl a ratio and its de-epoxidation state. The results indicate that NPQ can be used as a sensitive stress marker for nutrient deficiency, but cannot be used to estimate XCP pool size and state.

Xanthophyll cycle-related non-photochemical quenching protects Sargassum thunbergii from high light-induced photoinhibition

As a common macroalga living in the intertidal zone, Sargassum thunbergii (Sargassaceae, Phaeophyta) is often exposed to drastic changes in solar photosynthetically active radiation during a diurnal cycle; thus, the potential photosynthetic adaptation processes deserve attention. In this work, we examined the photosynthetic performance and xanthophyll cycle activity of this alga in response to high light (1,200 μmol photons m–2 s–1, the average in-situ light intensity at noon) by using chlorophyll fluorescence and high-performance liquid chromatography (HPLC). On exposure to high light, a rapid decrease in the effective quantum yield of photosystem II (PSII) (Y(II)) occurred, indicating down-regulation of PSII activity; a corresponding increase in non-photochemical quenching (NPQ) indicated the existence of energy-dissipating cycles. After turning off the light, Y(II) gradually increased to 0.7, accompanied by a decrease in NPQ. However, no complete recovery of NPQ was observed, and its value remained at ≈4, even after an 80-min dark treatment. The size of the xanthophyll cycle pigments pool was quantified using HPLC and was found to be ≈16 mol mol−1 Chl a × 100. The activity of the xanthophyll cycle, characterized by a de-epoxidation state (DPS), could reach up to ≈0.5. Such a large pigments pool and rapid accumulation of zeaxanthin may allow S. thunbergii to induce high values of NPQ (≈10). These results were further complemented by inhibitor (dithiothreitol, DTT) and pre-illumination experiments showing that (1) both NPQ and the xanthophyll cycle could be inhibited by DTT, and there was always a strong positive correlation between NPQ and DPS; (2) the previously formed antheraxanthin exhibited a long retention time and a slow epoxidation; and (3) the long retention of antheraxanthin contributed to a rapid accumulation of zeaxanthin. In conclusion, our present study demonstrated that xanthophyll cycle-induced NPQ can significantly protect S. thunbergii from light fluctuations in the intertidal zone.

Phenotyping of basil (Ocimum basilicum L.) illuminated with UV-A light of different wavelengths and intensities

The effects of light on plants depend on the energy dose and spectral characteristics. We investigated how UV-A radiation with two different wavelengths, 365 and 385 nm, combined at different intensities as supplemental light to red-blue baseline illumination affects growth and selected physiological and biochemical traits of green leaf basil (Ocimum basilicum L.). Conventional physiological measurements and biochemical analyses of photosynthetic pigments and phenolics were combined with high-throughput phenotyping. We did not detect any negative effects of UV-A on plant growth and photosynthetic functions, even at high doses of short-wave UV-A. Furthermore, thylakoid processes were enhanced under UV-A, as evidenced by higher electron transport rates and increased photochemical efficiency. The increase in anthocyanin index (ARI) indicates the UV-screening response. However, no anthocyanins were detected in the leaves, and the only phenolic compounds that responded to UV-A were hydroxycinnamic acid derivatives, which have a dual function in responding to UV-A (UV protection and antioxidants). UV-A increased the pool of xanthophyll cycle pigments and their ratio to chlorophylls. All this prevents photoinhibition and contributes to a high plasticity/efficiency of the basil's response to light environment.

Monitoring the photosynthetic performance of grape leaves using a hyperspectral-based machine learning model

Photosynthesis is a direct expression of the crop growth status and an important indicator predicting yield and quality. Rapid and accurate monitoring of the dynamics of photosynthetic is key to field management. In this study, we obtained photosynthetic pigments and water status parameters at the leaf scale during different growth periods of grape. The potential maximum quantum yield (Fv/Fm) of photosystem II (PSII) under dark adaptation and the light response curve (LRC) of the PSII electron transfer rate (ETR) under light adaptation were measured using a pulse amplitude modulated (PAM) chlorophyll fluorometer, while leaf spectral information was recorded using a hyperspectral imager. The maximum ETR (ETRmax) and initial quantum efficiency (kα) were calculated using the LRC model. A Bayesian neural network (BNN) model (implemented in Tensorfolw2.8) was developed to predict Fv/Fm, ETRmax and kα by quantifying the spectral response indices of photosynthetic pigments and water status parameters. A comparison was made with the partial least squares (PLS) and photochemical reflectance index (PRI). The results show that BNN, PLS model and PRI have better predictive performance for Fv/Fm than ETRmax and kα. Compared with the PLS and PRI, the BNN model was able to significantly improve the prediction accuracy, where the validation results for Fv/Fm were R2 of 0.78, ETRmax of 0.57 and kα of 0.53. In addition, the importance of the BNN model input features varied with Fv/Fm, ETRmax and kα, with the vegetation index describing the photosynthetic pigments having the highest importance. The PRI had the worst predictive performance probably because the de-epoxidation state of the xanthophyll cycle pigments is strongly influenced by temporal changes. The model developed in this paper for monitoring photosynthetic performance parameters in grape leaves can simplify the complex photosynthetic reaction process, expand the application of PAM technology and provide a method for rapid and accurate monitoring of photosynthetic performance.

Effects of Hypoxia on Coral Photobiology and Oxidative Stress.

Simple SummaryWith the growing importance of climate change research, the biological effects of oxygen loss on marine biota remain understudied. Coral reefs support diverse marine organisms and provide valuable ecosystem services. Deoxygenation may induce mass coral mortality and reduce species richness on coral reefs. Corals exposed to nocturnal hypoxia may experience detrimental effects due to induced oxidative stress caused by the increase in reactive oxygen species (ROS). The stress interferes with the most fundamental biological processes vital for the symbiosis between corals and photosynthetic algae. In this study, the coral Acropora spp. exhibited evident injury in photosynthetic apparatus, de-epoxidation state and DNA. However, besides that, there were no signs of engaged antioxidant defense mechanisms against ROS or pigment degradation, which leads to a conclusion that these corals may be resilient to such oxygen daily oscillations. Nevertheless, while corals might survive such short-term abiotic stress, the growing number and intensity of hypoxic events across the global ocean may pose a massive threat to these keystone invertebrate species.Global ocean oxygen (O2) content is decreasing as climate change drives declines in oxygen solubility, strengthened stratification of seawater masses, increased biological oxygen consumption and coastal eutrophication. Studies on the biological effects of nocturnal decreased oxygen concentrations (hypoxia) on coral reefs are very scarce. Coral reefs are fundamental for supporting one quarter of all marine species and essential for around 275 million people worldwide. This study investigates acute physiological and photobiological responses of a scleractinian coral (Acropora spp.) to overnight hypoxic conditions (<2 mg/L of O2). Bleaching was not detected, and visual and physical aspects of corals remained unchanged under hypoxic conditions. Most photobiological-related parameters also did not show significant changes between treatments. In addition to this, no significant differences between treatments were observed in the pigment composition. However, hypoxic conditions induced a significant decrease in coral de-epoxidation state of the xanthophyll cycle pigments and increase in DNA damage. Although the present findings suggest that Acropora spp. is resilient to some extent to short-term daily oxygen oscillations, long-term exposure to hypoxia, as predicted to occur with climate change, may still have deleterious effects on corals.

Photoacclimation impacts the molecular features of photosystem supercomplexes in the centric diatom Thalassiosira pseudonana

In diatoms, light-harvesting processes take place in a specific group of proteins, called fucoxanthin chlorophyll a/c proteins (FCP). This group includes many members and represents the major characteristic of the diatom photosynthetic apparatus, with specific pigments bound (chlorophyll c, fucoxanthin, diadino- and diatoxanthin besides chlorophyll a). In thylakoids, FCP and photosystems (PS) form multimeric supercomplexes.In this study, we compared the biochemical properties of PS supercomplexes isolated from Thalassiosira pseudonana cells grown under low light or high light conditions, respectively. High light acclimation changed the molecular features of the PS and their ratio in thylakoids. In PSII, no obvious changes in polypeptide composition were observed, whereas for PSI changes in one specific group of FCP proteins were detected. As reported before, the amount of xanthophyll cycle pigments and their de-epoxidation ratio was increased in PSI under HL. In PSII, however, no additional xanthophyll cycle pigments occurred, but the de-epoxidation ratio was increased as well. This comparison suggests how mechanisms of photoprotection might take place within and in the proximity of the PS, which gives new insights into the capacity of diatoms to adapt to different conditions and in different environments.

Haploid Helps Phaeocystis Globosa Distribute to Deeper Dim Water, as Evidenced by Growth and Photosynthetic Physiology

Phaeocystis globosa has a haplo-diplontic alternative life cycle and is an important causative species of harmful algal blooms. Diploid solitary cells of Phaeocystis can form colonies and bloom in the surface water. However, haploid cells are abundant in deep water rather than surface water. We hypothesize that the haploid cells of Phaeocystis globosa could better adapt to deep dim water than its diploid cells. Haploid and diploid solitary cells of P. globosa were cultured with eutrophic medium (f/2) under moderately low (15 µmol photons m-2 s-1), extremely low (5 µmol photons m-2 s-1) and normal (60 µmol photons m-2 s-1) irradiance conditions. The results showed that irradiances used in this study did not induce reactive oxygen species (ROS) damage in either haploid or diploid cells. Both haploid and diploid solitary cells grew faster at higher irradiance during the initial exponential growth phase. However, the haploid abundances under moderately and extremely low irradiances were higher than that under normal irradiance after the exponential growth phase, but diploid cells formed more colonies at higher irradiances. An increase in the photosynthetic pigments (PSC) ratio combined with a reduction in photoprotective pigments (PPC) ratio were found in both ploidies with decreasing irradiance, but the ratios of PSC and PPC and xanthophyll cycle pigments were significantly higher in haploid cells than in diploid cells. For haploids, the highest potential photochemistry efficiency of photosystems П was found under extremely low irradiance, but for diploids, it was observed under moderately low irradiance. The results suggest that both haploid and diploid solitary cells of P. globosa in eutrophic water can survive under low-light conditions, but haploid cells have an advantage in extremely low irradiance.

Isolation of fucoxanthin chlorophyll protein complexes of the centric diatom Thalassiosira pseudonana associated with the xanthophyll cycle enzyme diadinoxanthin de-epoxidase.

In the present study, low concentrations of the very mild detergent n-dodecyl-α-d-maltoside in conjunction with sucrose gradient ultracentrifugation were used to prepare fucoxanthin chlorophyll protein (FCP) complexes of the centric diatom Thalassiosira pseudonana. Two main FCP fractions were observed in the sucrose gradients, one in the upper part and one at high sucrose concentrations in the lower part of the gradient. The first fraction was dominated by the 18 kDa FCP protein band in SDS-gels. Since this fraction also contained other protein bands, it was designated as fraction enriched in FCP-A complex. The second fraction contained mainly the 21 kDa FCP band, which is typical for the FCP-B complex. Determination of the lipid composition showed that both FCP fractions contained monogalactosyl diacylglycerol as the main lipid followed by the second galactolipid of the thylakoid membrane, namely digalactosyl diacylglycerol. The negatively charged lipids sulfoquinovosyl diacylglycerol and phosphatidyl glycerol were also present in both fractions in pronounced concentrations. With respect to the pigment composition, the fraction enriched in FCP-A contained a higher amount of the xanthophyll cycle pigments diadinoxanthin (DD) and diatoxanthin (Dt), whereas the FCP-B fraction was characterized by a lower ratio of xanthophyll cycle pigments to the light-harvesting pigment fucoxanthin. Protein analysis by mass spectrometry revealed that in both FCP fractions the xanthophyll cycle enzyme diadinoxanthin de-epoxidase (DDE) was present. In addition, the analysis showed an enrichment of DDE in the fraction enriched in FCP-A but only a very low amount of DDE in the FCP-B fraction. In-vitro de-epoxidation assays, employing the isolated FCP complexes, were characterized by an inefficient conversion of DD to Dt. However, in line with the heterogeneous DDE distribution, the fraction enriched in FCP-A showed a more pronounced DD de-epoxidation compared with the FCP-B.

Relationship among light intensity, pigment compositions and photoinhibition in relation to rhodoxanthin accumulation in the gymnosperm Cryptomeria japonica

ABSTRACT Some gymnosperms, including Cryptomeria japonica, exhibit needle reddening caused by red xanthophyll pigment, rhodoxanthin accumulation during long-term cold acclimation. Although photoprotective role of rhodoxanthin, serves as a “sunscreen”, was proposed, the key environmental and/or physiological factors involved in rhodoxanthin accumulation remain unclear. We exposed C. japonica trees to four different light intensities (13%, 39%, 64% and 100% of full sunlight) and measured some parameters (the nocturnal de-epoxidation state [DPS: ratio of the zeaxanthin + antheraxanthin to xanthophyll cycle pigments], the β-carotene level and the maximum photosystem II efficiency [Fv/Fm]) that have been suggested to play roles in rhodoxanthin accumulation. Rhodoxanthin was synthesised from autumn to winter, and less so in spring. Light intensity related to the maximum value of rhodoxanthin on chlorophyll (Chl) basis, but not to that on FW basis, because Chl content decreased with increasing light intensity. These results indicated that rhodoxanthin accumulation was accompanied with Chl decrease. In addition, the relationship between the rhodoxanthin content and the seasonal growth temperature was different among light treatments, reflecting differences in the amount of excess light among light treatments Furthermore, the rhodoxanthin content correlated significantly with the Fv/Fm, nocturnal DPS and β-carotene level in winter. Thus, rhodoxanthin accumulates in a photoinhibitory environment occurs with a decrease in Chl content and an increase in DPS.

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.

Functional responses to climate change may increase invasive potential of Carpobrotus edulis.

Biological invasions and climate change are major threats to biodiversity. It is therefore important to anticipate how the climate changes projected for Southern Europe would affect the ecophysiological performance of the invasive South African plant, Carpobrotus edulis (ice plant or sour fig), and its capacity to undergo rapid adaptive evolution. We manipulated the climate conditions in a field plot located on the island of Sálvora (northwest of the Iberian Peninsula) to establish a full factorial experiment with C. edulis plants transplanted from four native (southern African) and four invasive (northwestern Iberian Peninsula) populations. Throughout 14 months we measured growth and functional traits of this species under two temperatures (control vs. increased), and two rainfall levels (control vs. reduced). Temperature increased photochemical efficiency and relative growth rate of C. edulis. Rainfall modulated some of the effects of temperature on C and N isotopic composition, and pigment contents. Invasive populations showed lower root mass allocation and higher survival rates, as well as increased water use efficiency, lipid peroxidation, chlorophyll, and xanthophyll cycle pigment contents than native populations. The increased growth and physiological performances observed under our experimental conditions suggest that the expected climate changes would further promote the invasion of C. edulis. Differences between native and invasive genotypes in survival and functional traits revealed that populations have diverged during the process of invasion, what gives support to the invasiveness hypothesis. Our findings highlight the importance of analyzing intraspecific variability in functional responses to better predict how invasive species will respond to environmental changes.

Drone-based physiological index reveals long-term acclimation and drought stress responses in trees.

Monitoring early tree physiological responses to drought is key to understanding progressive impacts of drought on forests and identifying resilient species. We combined drone‐based multispectral remote sensing with measurements of tree physiology and environmental parameters over two growing seasons in a 100‐y‐old Pinus sylvestris forest subject to 17‐y of precipitation manipulation. Our goal was to determine if drone‐based photochemical reflectance index (PRI) captures tree drought stress responses and whether responses are affected by long‐term acclimation. PRI detects changes in xanthophyll cycle pigment dynamics, which reflect increases in photoprotective non‐photochemical quenching activity resulting from drought‐induced photosynthesis downregulation. Here, PRI of never‐irrigated trees was up to 10 times lower (higher stress) than PRI of irrigated trees. Long‐term acclimation to experimental treatment, however, influenced the seasonal relationship between PRI and soil water availability. PRI also captured diurnal decreases in photochemical efficiency, driven by vapour pressure deficit. Interestingly, 5 years after irrigation was stopped for a subset of the irrigated trees, a positive legacy effect persisted, with lower stress responses (higher PRI) compared with never‐irrigated trees. This study demonstrates the ability of remotely sensed PRI to scale tree physiological responses to an entire forest and the importance of long‐term acclimation in determining current drought stress responses.

The Arabidopsis Accessions Selection Is Crucial: Insight from Photosynthetic Studies.

Natural genetic variation in photosynthesis is strictly associated with the remarkable adaptive plasticity observed amongst Arabidopsis thaliana accessions derived from environmentally distinct regions. Exploration of the characteristic features of the photosynthetic machinery could reveal the regulatory mechanisms underlying those traits. In this study, we performed a detailed characterisation and comparison of photosynthesis performance and spectral properties of the photosynthetic apparatus in the following selected Arabidopsis thaliana accessions commonly used in laboratories as background lines: Col-0, Col-1, Col-2, Col-8, Ler-0, and Ws-2. The main focus was to distinguish the characteristic disparities for every accession in photosynthetic efficiency that could be accountable for their remarkable plasticity to adapt. The biophysical and biochemical analysis of the thylakoid membranes in control conditions revealed differences in lipid-to-protein contribution, Chlorophyll-to-Carotenoid ratio (Chl/Car), and xanthophyll cycle pigment distribution among accessions. We presented that such changes led to disparities in the arrangement of the Chlorophyll-Protein complexes, the PSI/PSII ratio, and the lateral mobility of the thylakoid membrane, with the most significant aberrations detected in the Ler-0 and Ws-2 accessions. We concluded that selecting an accession suitable for specific research on the photosynthetic process is essential for optimising the experiment.

Fine-tuning of grapevine xanthophyll-cycle and energy dissipation under Mediterranean conditions by kaolin particle-film

Kaolin-particle film has been considered a low-cost technology to mitigate the adverse effects of high light and temperature, and drought in several crops. However, the underlying excess energy absorption and dissipation mechanisms, and related components associated with kaolin photoprotective effects in grapevines are poorly explored. This study aims to understand the interactions between kaolin foliar treatment and photosynthetic pigments accumulation, carotenoids metabolism, xanthophyll cycle regulation, and its putative role on the non-photochemical quenching (NPQ) processes in Touriga-Franca (TF) and Touriga-Nacional (TN) varieties. The experiments were conducted during the 2017 summer season in a commercial vineyard, and measurements were performed at pre-dawn and midday in each sampling date (EL35 – veraison; EL38 – full mature). Overall, TF variety showed higher accumulation of chlorophylls, xanthophylls, and de-epoxidation state (DPS) than TN. Kaolin treatment enhanced TN chlorophyll accumulation up to 114 % at EL35 (veraison) and 123 % at EL38 (full mature), highlighting its protective role on chlorophyll degradation, while no changes were found in TF, which might indicate a lower need for particle-film technology in this variety under the current environmental conditions. Individual carotenoids were mainly higher in the treated leaves of both varieties, as well as the xanthophyll cycle pigments zeaxanthin (Zx) and violaxanthin (Vx). Simultaneously, the DPS and NPQ values were lower in TN and TF treated leaves (1.92 – 2.36) compared to untreated vines (3.19 – 3.24), suggesting that there might be other components influencing NPQ levels beyond Zx, with an indirect role in long-lasting NPQ processes. In addition, in the TF kaolin-treated leaves, violaxanthin de-epoxidase (VvVDE1) and zeaxanthin epoxidase (VvZEP1) gene expression were respectively 3-fold and 4-fold upregulated at stage EL35, while VvZEP1 gene expression decreased at stage EL38 in TN kaolin-treated leaves, indicating an optimised regulation of the xanthophyll cycle. These findings suggest that kaolin treatment promoted a fine-tuning of grapevine summer stress responses under sustained summer stress factors, by managing xanthophyll cycle dynamics, and pigments accumulation.

In situ monitoring of chlorophyll a fluorescence in Nannochloropsis oceanica cultures to assess photochemical changes and the onset of lipid accumulation during nitrogen deprivation.

In situ chlorophyll a fluorescence measurements were applied to monitor changes in the photochemical variables of Nannochloropsis oceanica cultures under nitrogen-deplete and nitrogen-replete (control) conditions. In addition, growth, lipid, fatty acid, and pigment contents were also followed. In the control culture, growth was promoted along with pigment content, electron transport rate (ETR), and polyunsaturated fatty acids, while total lipid content and fatty acid saturation level diminished. Under nitrogen-deplete conditions, the culture showed a higher de-epoxidation state of the xanthophyll cycle pigments. Fast transients revealed a poor processing efficiency for electron transfer beyond QA , which was in line with the low ETR due to nitrogen depletion. Lipid content and the de-epoxidation state were the first biochemical variables triggered by the change in nutrient status, which coincided with a 20% drop in the in situ effective quantum yield of PSII (ΔF'/Fm '), and a raise in the Vj measurements. A good correlation was found between the changes in ΔF'/Fm ' and lipid content (r = -0.96, p < 0.01). The results confirm the reliability and applicability of in situ fluorescence measurements to monitor lipid induction in N. oceanica.

The fine-tuning of NPQ in diatoms relies on the regulation of both xanthophyll cycle enzymes

Diatoms possess an efficient mechanism to dissipate photons as heat in conditions of excess light, which is visualized as the Non-Photochemical Quenching of chlorophyll a fluorescence (NPQ). In most diatom species, NPQ is proportional to the concentration of the xanthophyll cycle pigment diatoxanthin formed from diadinoxanthin by the diadinoxanthin de-epoxidase enzyme. The reverse reaction is performed by the diatoxanthin epoxidase. Despite the xanthophyll cycle’s central role in photoprotection, its regulation is not yet well understood. The proportionality between diatoxanthin and NPQ allowed us to calculate the activity of both xanthophyll cycle enzymes in the model diatom Phaeodactylum tricornutum from NPQ kinetics. From there, we explored the light-dependency of the activity of both enzymes. Our results demonstrate that a tight regulation of both enzymes is key to fine-tune NPQ: (i) the rate constant of diadinoxanthin de-epoxidation is low under a light-limiting regime but increases as photosynthesis saturates, probably due to the thylakoidal proton gradient ΔpH (ii) the rate constant of diatoxanthin epoxidation exhibits an optimum under low light and decreases in the dark due to an insufficiency of the co-factor NADPH as well as in higher light through an as yet unresolved inhibition mechanism, that is unlikely to be related to the ΔpH. We observed that the suppression of NPQ by an uncoupler was due to an accelerated diatoxanthin epoxidation enzyme rather than to the usually hypothesized inhibition of the diadinoxanthin de-epoxidation enzyme.